[{"alternative_title":["ISTA Master’s Thesis"],"publication_identifier":{"issn":["2791-4585"]},"project":[{"_id":"8f347782-16d5-11f0-9cad-8c19706ee739","name":"Cyclic nucleotides as second messengers in plants","grant_number":"101142681"}],"publisher":"Institute of Science and Technology Austria","supervisor":[{"first_name":"Jiří","id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří"}],"oa_version":"Published Version","title":"Armadillo repeat only proteins are master regulators of plant cyclic-nucleotide gated channels","publication_status":"published","_id":"20964","year":"2026","article_processing_charge":"No","file":[{"embargo":"2027-01-01","file_name":"2026_Vladimirtsev_Dmitrii_Thesis.pdf","relation":"main_file","access_level":"closed","date_created":"2026-01-21T14:12:13Z","file_size":2867531,"content_type":"application/pdf","file_id":"21033","date_updated":"2026-01-21T14:12:13Z","checksum":"812857b2fbe3f6113bef22fd04bccd3e","creator":"dvladimi","embargo_to":"open_access"},{"file_name":"Source Files.zip","access_level":"closed","date_created":"2026-01-21T14:41:58Z","relation":"source_file","content_type":"application/x-zip-compressed","file_size":25023066,"file_id":"21034","date_updated":"2026-01-28T12:38:19Z","checksum":"2b969f97f8d7461bea3d255f48c2219c","creator":"dvladimi"}],"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT-ISTA-20964","citation":{"ieee":"D. Vladimirtsev, “Armadillo repeat only proteins are master regulators of plant cyclic-nucleotide gated channels,” Institute of Science and Technology Austria, 2026.","short":"D. Vladimirtsev, Armadillo Repeat Only Proteins Are Master Regulators of Plant Cyclic-Nucleotide Gated Channels, Institute of Science and Technology Austria, 2026.","ista":"Vladimirtsev D. 2026. Armadillo repeat only proteins are master regulators of plant cyclic-nucleotide gated channels. Institute of Science and Technology Austria.","chicago":"Vladimirtsev, Dmitrii. “Armadillo Repeat Only Proteins Are Master Regulators of Plant Cyclic-Nucleotide Gated Channels.” Institute of Science and Technology Austria, 2026. <a href=\"https://doi.org/10.15479/AT-ISTA-20964\">https://doi.org/10.15479/AT-ISTA-20964</a>.","mla":"Vladimirtsev, Dmitrii. <i>Armadillo Repeat Only Proteins Are Master Regulators of Plant Cyclic-Nucleotide Gated Channels</i>. Institute of Science and Technology Austria, 2026, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20964\">10.15479/AT-ISTA-20964</a>.","ama":"Vladimirtsev D. Armadillo repeat only proteins are master regulators of plant cyclic-nucleotide gated channels. 2026. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20964\">10.15479/AT-ISTA-20964</a>","apa":"Vladimirtsev, D. (2026). <i>Armadillo repeat only proteins are master regulators of plant cyclic-nucleotide gated channels</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-20964\">https://doi.org/10.15479/AT-ISTA-20964</a>"},"acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"Bio"}],"month":"01","date_published":"2026-01-14T00:00:00Z","ddc":["570"],"page":"22","status":"public","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"has_accepted_license":"1","type":"dissertation","degree_awarded":"MS","date_updated":"2026-04-07T11:41:44Z","OA_place":"publisher","related_material":{"record":[{"id":"20982","status":"public","relation":"part_of_dissertation"}]},"corr_author":"1","author":[{"full_name":"Vladimirtsev, Dmitrii","last_name":"Vladimirtsev","id":"60466724-5355-11ee-ae5a-fa55e8f99c3d","first_name":"Dmitrii"}],"file_date_updated":"2026-01-28T12:38:19Z","day":"14","language":[{"iso":"eng"}],"date_created":"2026-01-09T09:22:48Z"},{"license":"https://creativecommons.org/licenses/by/4.0/","publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publisher":"Institute of Science and Technology Austria","supervisor":[{"orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří","first_name":"Jiří","id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml"},{"first_name":"Eva","id":"38F4F166-F248-11E8-B48F-1D18A9856A87","last_name":"Benková","orcid":"0000-0002-8510-9739","full_name":"Benková, Eva"}],"title":"Mechanisms of auxin-mediated early embryogenesis in Arabidopsis thaliana","_id":"20362","publication_status":"published","oa_version":"Published Version","year":"2025","article_processing_charge":"No","file":[{"content_type":"application/pdf","file_size":7501548,"file_id":"20388","date_updated":"2025-09-26T07:29:11Z","creator":"dbabic","embargo_to":"open_access","checksum":"5ecf274281a54a41e0288bc79edf7492","embargo":"2026-09-25","file_name":"2025_David_Babic_Thesis.pdf","date_created":"2025-09-24T13:43:14Z","access_level":"closed","relation":"main_file"},{"file_name":"Thesis_Babic_draft.docx","access_level":"closed","date_created":"2025-09-24T13:43:14Z","relation":"source_file","file_size":23206052,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"20389","date_updated":"2025-09-26T07:29:11Z","creator":"dbabic","checksum":"2703e548390de0a1af7a707137e8ab3b"}],"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT-ISTA-20362","citation":{"ama":"Babic D. Mechanisms of auxin-mediated early embryogenesis in Arabidopsis thaliana. 2025. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20362\">10.15479/AT-ISTA-20362</a>","apa":"Babic, D. (2025). <i>Mechanisms of auxin-mediated early embryogenesis in Arabidopsis thaliana</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-20362\">https://doi.org/10.15479/AT-ISTA-20362</a>","mla":"Babic, David. <i>Mechanisms of Auxin-Mediated Early Embryogenesis in Arabidopsis Thaliana</i>. Institute of Science and Technology Austria, 2025, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20362\">10.15479/AT-ISTA-20362</a>.","chicago":"Babic, David. “Mechanisms of Auxin-Mediated Early Embryogenesis in Arabidopsis Thaliana.” Institute of Science and Technology Austria, 2025. <a href=\"https://doi.org/10.15479/AT-ISTA-20362\">https://doi.org/10.15479/AT-ISTA-20362</a>.","ista":"Babic D. 2025. Mechanisms of auxin-mediated early embryogenesis in Arabidopsis thaliana. Institute of Science and Technology Austria.","short":"D. Babic, Mechanisms of Auxin-Mediated Early Embryogenesis in Arabidopsis Thaliana, Institute of Science and Technology Austria, 2025.","ieee":"D. Babic, “Mechanisms of auxin-mediated early embryogenesis in Arabidopsis thaliana,” Institute of Science and Technology Austria, 2025."},"page":"116","status":"public","ddc":["580"],"date_published":"2025-09-18T00:00:00Z","acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"}],"month":"09","degree_awarded":"PhD","date_updated":"2026-04-07T11:52:02Z","department":[{"_id":"GradSch"},{"_id":"JiFr"},{"_id":"EvBe"}],"has_accepted_license":"1","type":"dissertation","corr_author":"1","OA_place":"publisher","related_material":{"record":[{"id":"20187","status":"public","relation":"part_of_dissertation"}]},"file_date_updated":"2025-09-26T07:29:11Z","date_created":"2025-09-17T13:28:01Z","day":"18","language":[{"iso":"eng"}],"author":[{"last_name":"Babic","id":"db566d23-f6e0-11ea-865d-e6f270e968e7","first_name":"David","full_name":"Babic, David"}]},{"keyword":["Auxin Signaling","Plant Development"],"year":"2025","title":"Nuclear and cell surface auxin signaling in A. thaliana developmental transitions","publication_status":"published","_id":"20364","oa_version":"Published Version","supervisor":[{"orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří","first_name":"Jiří","id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml"}],"publisher":"Institute of Science and Technology Austria","publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"file_date_updated":"2025-09-30T14:31:29Z","date_created":"2025-09-19T12:23:38Z","language":[{"iso":"eng"}],"day":"19","author":[{"id":"e3fdddd5-f6e0-11ea-865d-ca99ee6367f4","last_name":"Giannini","first_name":"Caterina","full_name":"Giannini, Caterina"}],"corr_author":"1","acknowledgement":"Plant Facility,\r\nProtein Service Facility","related_material":{"record":[{"relation":"part_of_dissertation","status":"public","id":"12291"},{"relation":"part_of_dissertation","status":"public","id":"19399"}]},"OA_place":"publisher","degree_awarded":"PhD","date_updated":"2026-07-06T12:51:13Z","has_accepted_license":"1","department":[{"_id":"GradSch"},{"_id":"JiFr"},{"_id":"MaLo"}],"type":"dissertation","page":"151","status":"public","ddc":["580"],"month":"09","date_published":"2025-09-19T00:00:00Z","acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"Bio"}],"citation":{"ieee":"C. Giannini, “Nuclear and cell surface auxin signaling in A. thaliana developmental transitions,” Institute of Science and Technology Austria, 2025.","ista":"Giannini C. 2025. Nuclear and cell surface auxin signaling in A. thaliana developmental transitions. Institute of Science and Technology Austria.","short":"C. Giannini, Nuclear and Cell Surface Auxin Signaling in A. Thaliana Developmental Transitions, Institute of Science and Technology Austria, 2025.","chicago":"Giannini, Caterina. “Nuclear and Cell Surface Auxin Signaling in A. Thaliana Developmental Transitions.” Institute of Science and Technology Austria, 2025. <a href=\"https://doi.org/10.15479/AT-ISTA-20364\">https://doi.org/10.15479/AT-ISTA-20364</a>.","mla":"Giannini, Caterina. <i>Nuclear and Cell Surface Auxin Signaling in A. Thaliana Developmental Transitions</i>. Institute of Science and Technology Austria, 2025, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20364\">10.15479/AT-ISTA-20364</a>.","ama":"Giannini C. Nuclear and cell surface auxin signaling in A. thaliana developmental transitions. 2025. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20364\">10.15479/AT-ISTA-20364</a>","apa":"Giannini, C. (2025). <i>Nuclear and cell surface auxin signaling in A. thaliana developmental transitions</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-20364\">https://doi.org/10.15479/AT-ISTA-20364</a>"},"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT-ISTA-20364","article_processing_charge":"No","file":[{"relation":"main_file","access_level":"closed","date_created":"2025-09-24T14:46:34Z","file_name":"2025_Giannini_Caterina_Thesis...pdf","embargo":"2026-09-30","checksum":"536ba1701453b0b2346be14c046b2911","embargo_to":"open_access","creator":"cgiannin","date_updated":"2025-09-30T14:31:29Z","file_id":"20390","file_size":14278965,"content_type":"application/pdf"},{"relation":"source_file","date_created":"2025-09-24T14:46:35Z","access_level":"closed","file_name":"2025_Giannini_Caterina_Thesis...docx","date_updated":"2025-09-24T14:46:35Z","creator":"cgiannin","checksum":"192f55262f2da2ea0a59d9c23a1b8573","content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_size":24499022,"file_id":"20391"}]},{"year":"2025","ec_funded":1,"oa_version":"Published Version","title":"The cAMP second messenger in auxin signalling","_id":"19478","publication_status":"published","publisher":"Institute of Science and Technology Austria","supervisor":[{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"}],"alternative_title":["ISTA Thesis"],"publication_identifier":{"issn":["2663-337X"]},"project":[{"name":"Tracing Evolution of Auxin Transport and Polarity in Plants","_id":"261099A6-B435-11E9-9278-68D0E5697425","grant_number":"742985","call_identifier":"H2020"},{"name":"Guanylate cyclase activity of TIR1/AFBs auxin receptors","_id":"7bcece63-9f16-11ee-852c-ae94e099eeb6","grant_number":"P37051"},{"name":"Cyclic nucleotides as second messengers in plants","_id":"8f347782-16d5-11f0-9cad-8c19706ee739","grant_number":"101142681"}],"OA_place":"publisher","related_material":{"record":[{"status":"public","id":"19421","relation":"part_of_dissertation"},{"id":"13212","status":"public","relation":"part_of_dissertation"}]},"corr_author":"1","acknowledgement":"This project was funded by the European Research Council Advanced Grant (ETAP-742985),\r\nEuropean Research Council (ERC; 101142681 CYNIPS), Austrian Science Fund (FWF; P\r\n37051-B).","author":[{"first_name":"Huihuang","id":"83c96512-15b2-11ec-abd3-b7eede36184f","last_name":"Chen","full_name":"Chen, Huihuang"}],"file_date_updated":"2025-04-09T13:53:38Z","date_created":"2025-04-04T07:48:24Z","language":[{"iso":"eng"}],"day":"04","date_published":"2025-04-04T00:00:00Z","ddc":["580"],"acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"}],"month":"04","page":"118","status":"public","has_accepted_license":"1","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"type":"dissertation","degree_awarded":"PhD","date_updated":"2026-07-06T12:58:58Z","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT-ISTA-19478","citation":{"ieee":"H. Chen, “The cAMP second messenger in auxin signalling,” Institute of Science and Technology Austria, 2025.","short":"H. Chen, The CAMP Second Messenger in Auxin Signalling, Institute of Science and Technology Austria, 2025.","ista":"Chen H. 2025. The cAMP second messenger in auxin signalling. Institute of Science and Technology Austria.","chicago":"Chen, Huihuang. “The CAMP Second Messenger in Auxin Signalling.” Institute of Science and Technology Austria, 2025. <a href=\"https://doi.org/10.15479/AT-ISTA-19478\">https://doi.org/10.15479/AT-ISTA-19478</a>.","ama":"Chen H. The cAMP second messenger in auxin signalling. 2025. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-19478\">10.15479/AT-ISTA-19478</a>","apa":"Chen, H. (2025). <i>The cAMP second messenger in auxin signalling</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-19478\">https://doi.org/10.15479/AT-ISTA-19478</a>","mla":"Chen, Huihuang. <i>The CAMP Second Messenger in Auxin Signalling</i>. Institute of Science and Technology Austria, 2025, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-19478\">10.15479/AT-ISTA-19478</a>."},"article_processing_charge":"No","file":[{"date_updated":"2025-04-08T08:22:37Z","creator":"hchen","checksum":"b154973663a1bba505683faab7ae5ead","content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_size":16344814,"file_id":"19526","relation":"source_file","access_level":"closed","date_created":"2025-04-08T08:00:07Z","file_name":"Thesis_0403_Huihuang.docx"},{"file_name":"Thesis_0406_PDFA_Huihuang_1.pdf","embargo":"2026-10-08","access_level":"closed","date_created":"2025-04-08T08:00:06Z","relation":"main_file","file_id":"19527","file_size":8482147,"content_type":"application/pdf","creator":"hchen","checksum":"0099565f024388830c125ec17375c1a0","embargo_to":"local","date_updated":"2025-04-09T13:53:38Z"}]},{"author":[{"full_name":"Monzer, Aline","first_name":"Aline","id":"2DB5D88C-D7B3-11E9-B8FD-7907E6697425","last_name":"Monzer"}],"file_date_updated":"2025-04-01T07:55:27Z","date_created":"2025-03-12T14:25:42Z","language":[{"iso":"eng"}],"day":"13","OA_place":"publisher","related_material":{"record":[{"status":"public","id":"12291","relation":"part_of_dissertation"},{"id":"14826","status":"public","relation":"part_of_dissertation"},{"id":"19399","status":"public","relation":"part_of_dissertation"},{"id":"19398","status":"public","relation":"part_of_dissertation"}]},"corr_author":"1","acknowledgement":"I would like to acknowledge the facilities at ISTA, particularly LSF, IOF, and, of course, the plant facility, for providing the necessary resources for my research.","doi_confirm":"1","has_accepted_license":"1","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"type":"dissertation","degree_awarded":"PhD","date_updated":"2026-08-14T09:33:46Z","month":"03","date_published":"2025-03-13T00:00:00Z","ddc":["580"],"acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"Bio"}],"page":"160","status":"public","citation":{"short":"A. Monzer, Cell-Surface Auxin Signaling: Linking Molecular Pathways to Plant Development, Institute of Science and Technology Austria, 2025.","ista":"Monzer A. 2025. Cell-surface auxin signaling: Linking molecular pathways to plant development. Institute of Science and Technology Austria.","ieee":"A. Monzer, “Cell-surface auxin signaling: Linking molecular pathways to plant development,” Institute of Science and Technology Austria, 2025.","apa":"Monzer, A. (2025). <i>Cell-surface auxin signaling: Linking molecular pathways to plant development</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-19395\">https://doi.org/10.15479/AT-ISTA-19395</a>","ama":"Monzer A. Cell-surface auxin signaling: Linking molecular pathways to plant development. 2025. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-19395\">10.15479/AT-ISTA-19395</a>","mla":"Monzer, Aline. <i>Cell-Surface Auxin Signaling: Linking Molecular Pathways to Plant Development</i>. Institute of Science and Technology Austria, 2025, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-19395\">10.15479/AT-ISTA-19395</a>.","chicago":"Monzer, Aline. “Cell-Surface Auxin Signaling: Linking Molecular Pathways to Plant Development.” Institute of Science and Technology Austria, 2025. <a href=\"https://doi.org/10.15479/AT-ISTA-19395\">https://doi.org/10.15479/AT-ISTA-19395</a>."},"doi":"10.15479/AT-ISTA-19395","user_id":"8b945eb4-e2f2-11eb-945a-df72226e66a9","article_processing_charge":"No","file":[{"success":1,"file_name":"Final Thesis Aline Monzer.pdf","relation":"main_file","access_level":"open_access","date_created":"2025-03-12T14:14:49Z","file_id":"19396","file_size":13119670,"content_type":"application/pdf","creator":"amonzer","checksum":"9a3dd03bb4ec6b9907a325c3c4e8a1d7","date_updated":"2025-03-12T14:14:49Z"},{"date_updated":"2025-04-01T07:55:27Z","checksum":"a353ce1ee2eabce37bca35499e76dbf1","creator":"amonzer","file_size":13774837,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"19397","access_level":"closed","date_created":"2025-03-12T14:15:19Z","relation":"source_file","file_name":"Thesis Aline.docx"}],"abstract":[{"lang":"eng","text":"Plant growth and development rely significantly on phytohormones, with auxin serving as a master regulator, orchestrating processes from embryogenesis to organogenesis, vascular patterning, and environmental adaptation. Since its conceptual proposition by Charles Darwin in 1880 as an endogenous chemical signal influencing phototropism in grass, auxin has captivated scientists seeking to understand how such a small molecule exerts a profound influence on plant development.\r\nOne particularly fascinating aspect of auxin function is its ability to self-organize its transport. Through a feedback mechanism between auxin perception and directional transport—primarily mediated by PIN auxin transporters—auxin establishes narrow transport channels. This phenomenon, known as auxin canalization, is fundamental to vascular formation, regeneration, and other key developmental processes. Despite advances in our understanding, driven by experimental studies and computational models, auxin canalization remains an enigma, with many unanswered questions.\r\nLike other hormones, auxin functions through intricate signaling pathways. It operates through at least two distinct signaling mechanisms: the well-characterized canonical pathway and the less understood non-canonical pathway. While significant progress has been made in elucidating the canonical pathway, the non-canonical mechanisms remain less defined and require further investigation.\r\nIn this study, we revisit the non-canonical auxin signaling pathway mediated by the cell-surface complex Auxin Binding Protein 1-Transmembrane Kinase 1 (ABP1-TMK1), with a particular focus on its downstream phosphorylation events. We reveal that this auxin-mediated phosphorylation is conserved across the green lineage, underscoring its fundamental role in plant development. We explore key phosphorylation targets, particularly PIN2, which is essential for root gravitropism. To further understand TMK1’s role in diverse developmental processes, we identified and investigated its interactors as potential co-receptors or regulatory components within its signaling network.\r\nGiven the previously established role of ABP1-TMK1 in auxin canalization, we sought to further investigate this process and identified several TMK1 interactors also involved in this intricate mechanism.\r\nThese findings provide new insights into the complex regulation of auxin canalization, highlighting a broader and more interconnected signaling framework than previously understood."}],"year":"2025","oa_version":"Published Version","title":"Cell-surface auxin signaling: Linking molecular pathways to plant development","_id":"19395","publication_status":"published","oa":1,"supervisor":[{"full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596","id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří"}],"publisher":"Institute of Science and Technology Austria","alternative_title":["ISTA Thesis"],"publication_identifier":{"eisbn":["978-3-99078-054-1"],"eissn":["2663-337X"]}},{"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/at:ista:14510","citation":{"short":"N. Gnyliukh, Mechanism of Clathrin-Coated Vesicle  Formation during Endocytosis in Plants, Institute of Science and Technology Austria, 2023.","ista":"Gnyliukh N. 2023. Mechanism of clathrin-coated vesicle  formation during endocytosis in plants. Institute of Science and Technology Austria.","ieee":"N. Gnyliukh, “Mechanism of clathrin-coated vesicle  formation during endocytosis in plants,” Institute of Science and Technology Austria, 2023.","apa":"Gnyliukh, N. (2023). <i>Mechanism of clathrin-coated vesicle  formation during endocytosis in plants</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/at:ista:14510\">https://doi.org/10.15479/at:ista:14510</a>","ama":"Gnyliukh N. Mechanism of clathrin-coated vesicle  formation during endocytosis in plants. 2023. doi:<a href=\"https://doi.org/10.15479/at:ista:14510\">10.15479/at:ista:14510</a>","mla":"Gnyliukh, Nataliia. <i>Mechanism of Clathrin-Coated Vesicle  Formation during Endocytosis in Plants</i>. Institute of Science and Technology Austria, 2023, doi:<a href=\"https://doi.org/10.15479/at:ista:14510\">10.15479/at:ista:14510</a>.","chicago":"Gnyliukh, Nataliia. “Mechanism of Clathrin-Coated Vesicle  Formation during Endocytosis in Plants.” Institute of Science and Technology Austria, 2023. <a href=\"https://doi.org/10.15479/at:ista:14510\">https://doi.org/10.15479/at:ista:14510</a>."},"file":[{"embargo_to":"open_access","creator":"ngnyliuk","checksum":"3d5e680bfc61f98e308c434f45cc9bd6","date_updated":"2024-11-23T23:30:38Z","file_id":"14567","file_size":20824903,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","date_created":"2023-11-20T09:18:51Z","access_level":"closed","relation":"source_file","file_name":"Thesis_Gnyliukh_final_08_11_23.docx"},{"date_updated":"2024-11-23T23:30:38Z","creator":"ngnyliuk","checksum":"bfc96d47fc4e7e857dd71656097214a4","file_size":24871844,"content_type":"application/pdf","file_id":"14568","date_created":"2023-11-20T09:23:11Z","access_level":"open_access","relation":"main_file","embargo":"2024-11-23","file_name":"Thesis_Gnyliukh_final_20_11_23.pdf"}],"article_processing_charge":"No","OA_place":"publisher","related_material":{"record":[{"status":"public","id":"14591","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","status":"public","id":"9887"},{"relation":"part_of_dissertation","status":"public","id":"8139"}]},"corr_author":"1","author":[{"full_name":"Gnyliukh, Nataliia","orcid":"0000-0002-2198-0509","id":"390C1120-F248-11E8-B48F-1D18A9856A87","last_name":"Gnyliukh","first_name":"Nataliia"}],"language":[{"iso":"eng"}],"day":"10","date_created":"2023-11-10T09:10:06Z","file_date_updated":"2024-11-23T23:30:38Z","acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"Bio"},{"_id":"LifeSc"}],"date_published":"2023-11-10T00:00:00Z","month":"11","ddc":["570"],"status":"public","page":"180","type":"dissertation","department":[{"_id":"GradSch"},{"_id":"JiFr"},{"_id":"MaLo"}],"has_accepted_license":"1","date_updated":"2026-08-24T22:30:57Z","degree_awarded":"PhD","publisher":"Institute of Science and Technology Austria","supervisor":[{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"},{"full_name":"Loose, Martin","orcid":"0000-0001-7309-9724","id":"462D4284-F248-11E8-B48F-1D18A9856A87","last_name":"Loose","first_name":"Martin"}],"alternative_title":["ISTA Thesis"],"publication_identifier":{"isbn":["978-3-99078-037-4"],"issn":["2663-337X"]},"project":[{"name":"International IST Doctoral Program","_id":"2564DBCA-B435-11E9-9278-68D0E5697425","grant_number":"665385","call_identifier":"H2020"}],"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"year":"2023","abstract":[{"text":"Clathrin-mediated endocytosis (CME) is vital for the regulation of plant growth and\r\ndevelopment by controlling plasma membrane protein composition and cargo uptake. CME\r\nrelies on the precise recruitment control of protein regulators for vesicle maturation and\r\nrelease. During the early stages of endocytosis, an area of flat membrane is remodelled by\r\nproteins to create a spherical vesicle against intracellular forces. After the Clathrin-coated\r\nvesicle (CCV) is fully formed, scission machinery releases it from the plasma membrane,\r\nand cargo proceeds for recycling or degradation through early endosomes / Trans Golgi\r\nnetwork. Protein machineries that mediate membrane bending and vesicle release in plants\r\nare unknown. However, studies show, that plant endocytosis is actin independent, thus\r\nindicating that plants utilize a unique mechanism to mediate membrane bending against highturgor pressure compared to other model systems. First, by using biochemical and advanced\r\nlive microscopy approaches we investigate the TPLATE complex, a plant-specific\r\nendocytosis protein complex. We found that TPLATE is peripherally associated with\r\nclathrin-coated vesicles and localises at the rim of endocytosis events. Next, our study of\r\nplant Dynamin-related protein 1C (DRP1C), which was hypothesised previously to play a\r\nrole in vesicle release, shows the recruitment of the protein already at the early stages of\r\nendocytosis. Moreover, DRP1C assembles into organised ring-like structures and is able to\r\ninduce membrane deformation and tubulation, suggesting its role also in membrane bending\r\nduring early CME. Based on the data from mammalian and yeast systems, plant DynaminRelated Proteins 2 and SH3P2 protein are strong candidates to be part of the plant vesicle\r\nscission machinery; however, their precise role in plant CME has not been yet elucidated.\r\nHere, we characterised DRP2s and SH3P2 roles in CME by combining high-resolution\r\nimaging of endocytic events in vivo and protein characterisation. Although DRP2s and\r\nSH3P2 arrive together during late CME and physically interact, genetic analysis using\r\n∆sh3p1,2,3 mutant and complementation with non-DRP2-interacting SH3P2 variants suggest\r\nthat SH3P2 does not directly recruit DRP2s to the site of endocytosis. Summarising our\r\nresearch, these observations provide new important insights into the mechanism of plant\r\nCME and show that, despite plants posses many homologues of mammalian and yeast CME\r\ncomponents, they do not necessarily act in the same manner. ","lang":"eng"}],"keyword":["Clathrin-Mediated Endocytosis","vesicle scission","Dynamin-Related Protein 2","SH3P2","TPLATE complex","Total internal reflection fluorescence microscopy","Arabidopsis thaliana"],"ec_funded":1,"oa":1,"oa_version":"Published Version","publication_status":"published","_id":"14510","title":"Mechanism of clathrin-coated vesicle  formation during endocytosis in plants"},{"publisher":"Institute of Science and Technology Austria","supervisor":[{"last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"},{"first_name":"Eva","last_name":"Benková","id":"38F4F166-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8510-9739","full_name":"Benková, Eva"},{"full_name":"Shani, Eilon","last_name":"Shani","first_name":"Eilon"}],"publication_identifier":{"isbn":["978-3-99078-019-0"],"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"project":[{"name":"Tracing Evolution of Auxin Transport and Polarity in Plants","_id":"261099A6-B435-11E9-9278-68D0E5697425","call_identifier":"H2020","grant_number":"742985"}],"year":"2022","abstract":[{"text":"Plant growth and development is well known to be both, flexible and dynamic. The high capacity for post-embryonic organ formation and tissue regeneration requires tightly regulated intercellular communication and coordinated tissue polarization. One of the most important drivers for patterning and polarity in plant development is the phytohormone auxin. Auxin has the unique characteristic to establish polarized channels for its own active directional cell to cell transport. This fascinating phenomenon is called auxin canalization. Those auxin transport channels are characterized by the expression and polar, subcellular localization of PIN auxin efflux carriers. PIN proteins have the ability to dynamically change their localization and auxin itself can affect this by interfering with trafficking. Most of the underlying molecular mechanisms of canalization still remain enigmatic. What is known so far is that canonical auxin signaling is indispensable but also other non-canonical signaling components are thought to play a role. In order to shed light into the mysteries auf auxin canalization this study revisits the branches of auxin signaling in detail. Further a new auxin analogue, PISA, is developed which triggers auxin-like responses but does not directly activate canonical transcriptional auxin signaling. We revisit the direct auxin effect on PIN trafficking where we found that, contradictory to previous observations, auxin is very specifically promoting endocytosis of PIN2 but has no overall effect on endocytosis. Further, we evaluate which cellular processes related to PIN subcellular dynamics are involved in the establishment of auxin conducting channels and the formation of vascular tissue. We are re-evaluating the function of AUXIN BINDING PROTEIN 1 (ABP1) and provide a comprehensive picture about its developmental phneotypes and involvement in auxin signaling and canalization. Lastly, we are focusing on the crosstalk between the hormone strigolactone (SL) and auxin and found that SL is interfering with essentially all processes involved in auxin canalization in a non-transcriptional manner. Lastly we identify a new way of SL perception and signaling which is emanating from mitochondria, is independent of canonical SL signaling and is modulating primary root growth.","lang":"eng"}],"ec_funded":1,"oa":1,"_id":"11626","publication_status":"published","title":"Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana","oa_version":"Published Version","doi":"10.15479/at:ista:11626","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","citation":{"ista":"Gallei MC. 2022. Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana. Institute of Science and Technology Austria.","short":"M.C. Gallei, Auxin and Strigolactone Non-Canonical Signaling Regulating Development in Arabidopsis Thaliana, Institute of Science and Technology Austria, 2022.","ieee":"M. C. Gallei, “Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana,” Institute of Science and Technology Austria, 2022.","apa":"Gallei, M. C. (2022). <i>Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/at:ista:11626\">https://doi.org/10.15479/at:ista:11626</a>","ama":"Gallei MC. Auxin and strigolactone non-canonical signaling regulating development in Arabidopsis thaliana. 2022. doi:<a href=\"https://doi.org/10.15479/at:ista:11626\">10.15479/at:ista:11626</a>","mla":"Gallei, Michelle C. <i>Auxin and Strigolactone Non-Canonical Signaling Regulating Development in Arabidopsis Thaliana</i>. Institute of Science and Technology Austria, 2022, doi:<a href=\"https://doi.org/10.15479/at:ista:11626\">10.15479/at:ista:11626</a>.","chicago":"Gallei, Michelle C. “Auxin and Strigolactone Non-Canonical Signaling Regulating Development in Arabidopsis Thaliana.” Institute of Science and Technology Austria, 2022. <a href=\"https://doi.org/10.15479/at:ista:11626\">https://doi.org/10.15479/at:ista:11626</a>."},"file":[{"checksum":"bd7ac35403cf5b4b2607287d2a104b3a","creator":"mgallei","date_updated":"2022-07-25T09:08:47Z","file_id":"11645","content_type":"application/pdf","file_size":9730864,"date_created":"2022-07-25T09:08:47Z","access_level":"open_access","relation":"main_file","file_name":"Thesis_Gallei.pdf"},{"file_size":19560720,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"11646","date_updated":"2022-07-25T09:39:58Z","creator":"mgallei","checksum":"a9e54fe5471ba25dc13c2150c1b8ccbb","file_name":"Thesis_Gallei_source.docx","relation":"source_file","access_level":"closed","date_created":"2022-07-25T09:09:09Z"},{"relation":"source_file","date_created":"2022-07-25T09:09:32Z","access_level":"closed","description":"This is the print version of the thesis including the full appendix","file_name":"Thesis_Gallei_to_print.pdf","date_updated":"2022-07-25T09:39:58Z","checksum":"3994f7f20058941b5bb8a16886b21e71","creator":"mgallei","file_size":24542837,"content_type":"application/pdf","file_id":"11647"},{"file_name":"Thesis_Gallei_Appendix.pdf","date_created":"2022-07-25T11:48:45Z","access_level":"open_access","relation":"main_file","file_size":15435966,"content_type":"application/pdf","file_id":"11650","date_updated":"2022-07-25T11:48:45Z","creator":"mgallei","checksum":"f24acd3c0d864f4c6676e8b0d7bfa76b"}],"article_processing_charge":"No","corr_author":"1","OA_place":"publisher","related_material":{"record":[{"status":"public","id":"8138","relation":"part_of_dissertation"},{"id":"7142","status":"public","relation":"part_of_dissertation"},{"status":"public","id":"10411","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","status":"public","id":"8931"},{"relation":"part_of_dissertation","status":"public","id":"7465"},{"status":"public","id":"9287","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","status":"public","id":"6260"}]},"date_created":"2022-07-20T11:21:53Z","language":[{"iso":"eng"}],"day":"20","file_date_updated":"2022-07-25T11:48:45Z","author":[{"first_name":"Michelle C","id":"35A03822-F248-11E8-B48F-1D18A9856A87","last_name":"Gallei","orcid":"0000-0003-1286-7368","full_name":"Gallei, Michelle C"}],"status":"public","page":"248","date_published":"2022-07-20T00:00:00Z","month":"07","ddc":["575"],"date_updated":"2026-06-18T19:02:05Z","degree_awarded":"PhD","type":"dissertation","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"has_accepted_license":"1"},{"abstract":[{"lang":"eng","text":"Blood – this is what animals use to heal wounds fast and efficient. Plants do not have blood circulation and their cells cannot move. However, plants have evolved remarkable capacities to regenerate tissues and organs preventing further damage. In my PhD research, I studied the wound healing in the Arabidopsis root. I used a UV laser to ablate single cells in the root tip and observed the consequent wound healing. Interestingly, the inner adjacent cells induced a\r\ndivision plane switch and subsequently adopted the cell type of the killed cell to replace it. We termed this form of wound healing “restorative divisions”. This initial observation triggered the questions of my PhD studies: How and why do cells orient their division planes, how do they feel the wound and why does this happen only in inner adjacent cells.\r\nFor answering these questions, I used a quite simple experimental setup: 5 day - old seedlings were stained with propidium iodide to visualize cell walls and dead cells; ablation was carried out using a special laser cutter and a confocal microscope. Adaptation of the novel vertical microscope system made it possible to observe wounds in real time. This revealed that restorative divisions occur at increased frequency compared to normal divisions. Additionally,\r\nthe major plant hormone auxin accumulates in wound adjacent cells and drives the expression of the wound-stress responsive transcription factor ERF115. Using this as a marker gene for wound responses, we found that an important part of wound signalling is the sensing of the collapse of the ablated cell. The collapse causes a radical pressure drop, which results in strong tissue deformations. These deformations manifest in an invasion of the now free spot specifically by the inner adjacent cells within seconds, probably because of higher pressure of the inner tissues. Long-term imaging revealed that those deformed cells continuously expand towards the wound hole and that this is crucial for the restorative division. These wound-expanding cells exhibit an abnormal, biphasic polarity of microtubule arrays\r\nbefore the division. Experiments inhibiting cell expansion suggest that it is the biphasic stretching that induces those MT arrays. Adapting the micromanipulator aspiration system from animal scientists at our institute confirmed the hypothesis that stretching influences microtubule stability. In conclusion, this shows that microtubules react to tissue deformation\r\nand this facilitates the observed division plane switch. This puts mechanical cues and tensions at the most prominent position for explaining the growth and wound healing properties of plants. Hence, it shines light onto the importance of understanding mechanical signal transduction. "}],"year":"2021","title":"Wound healing in the Arabidopsis root meristem","_id":"9992","publication_status":"published","oa_version":"Published Version","ec_funded":1,"oa":1,"supervisor":[{"orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří","first_name":"Jiří","last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87"}],"publisher":"Institute of Science and Technology Austria","tmp":{"short":"CC BY-NC-ND (4.0)","image":"/images/cc_by_nc_nd.png","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode"},"project":[{"name":"RNA-directed DNA methylation in plant development","_id":"262EF96E-B435-11E9-9278-68D0E5697425","grant_number":"P29988","call_identifier":"FWF"},{"name":"Tracing Evolution of Auxin Transport and Polarity in Plants","_id":"261099A6-B435-11E9-9278-68D0E5697425","grant_number":"742985","call_identifier":"H2020"}],"license":"https://creativecommons.org/licenses/by-nc-nd/4.0/","publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"file_date_updated":"2021-09-15T22:30:26Z","date_created":"2021-09-09T07:37:20Z","language":[{"iso":"eng"}],"day":"13","author":[{"first_name":"Lukas","last_name":"Hörmayer","id":"2EEE7A2A-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8295-2926","full_name":"Hörmayer, Lukas"}],"corr_author":"1","related_material":{"record":[{"status":"public","id":"6943","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","id":"8002","status":"public"},{"status":"public","id":"6351","relation":"part_of_dissertation"}]},"OA_place":"publisher","degree_awarded":"PhD","date_updated":"2026-04-08T07:11:47Z","has_accepted_license":"1","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"type":"dissertation","page":"168","status":"public","ddc":["575"],"acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"}],"month":"09","date_published":"2021-09-13T00:00:00Z","citation":{"ieee":"L. Hörmayer, “Wound healing in the Arabidopsis root meristem,” Institute of Science and Technology Austria, 2021.","ista":"Hörmayer L. 2021. Wound healing in the Arabidopsis root meristem. Institute of Science and Technology Austria.","short":"L. Hörmayer, Wound Healing in the Arabidopsis Root Meristem, Institute of Science and Technology Austria, 2021.","chicago":"Hörmayer, Lukas. “Wound Healing in the Arabidopsis Root Meristem.” Institute of Science and Technology Austria, 2021. <a href=\"https://doi.org/10.15479/at:ista:9992\">https://doi.org/10.15479/at:ista:9992</a>.","apa":"Hörmayer, L. (2021). <i>Wound healing in the Arabidopsis root meristem</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/at:ista:9992\">https://doi.org/10.15479/at:ista:9992</a>","ama":"Hörmayer L. Wound healing in the Arabidopsis root meristem. 2021. doi:<a href=\"https://doi.org/10.15479/at:ista:9992\">10.15479/at:ista:9992</a>","mla":"Hörmayer, Lukas. <i>Wound Healing in the Arabidopsis Root Meristem</i>. Institute of Science and Technology Austria, 2021, doi:<a href=\"https://doi.org/10.15479/at:ista:9992\">10.15479/at:ista:9992</a>."},"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/at:ista:9992","article_processing_charge":"No","file":[{"file_size":25179004,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"9993","date_updated":"2021-09-15T22:30:26Z","embargo_to":"open_access","creator":"lhoermaye","checksum":"c763064adaa720e16066c1a4f9682bbb","file_name":"Thesis_vupload.docx","relation":"source_file","date_created":"2021-09-09T07:29:48Z","access_level":"closed"},{"content_type":"application/pdf","file_size":6246900,"file_id":"9996","date_updated":"2021-09-15T22:30:26Z","creator":"lhoermaye","checksum":"53911b06e93d7cdbbf4c7f4c162fa70f","embargo":"2021-09-09","file_name":"Thesis_vfinal_pdfa.pdf","relation":"main_file","date_created":"2021-09-09T14:25:08Z","access_level":"open_access"}]},{"author":[{"first_name":"Lanxin","last_name":"Li","id":"367EF8FA-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-5607-272X","full_name":"Li, Lanxin"}],"date_created":"2021-10-04T13:33:10Z","language":[{"iso":"eng"}],"day":"06","file_date_updated":"2022-12-20T23:30:03Z","OA_place":"publisher","related_material":{"record":[{"relation":"part_of_dissertation","id":"442","status":"public"},{"relation":"part_of_dissertation","id":"6627","status":"public"},{"relation":"part_of_dissertation","id":"8931","status":"public"},{"relation":"part_of_dissertation","id":"8986","status":"public"},{"relation":"part_of_dissertation","id":"10095","status":"public"},{"relation":"part_of_dissertation","id":"8283","status":"public"},{"id":"9287","status":"public","relation":"part_of_dissertation"},{"status":"public","id":"10015","relation":"part_of_dissertation"}]},"corr_author":"1","type":"dissertation","department":[{"_id":"GradSch"},{"_id":"JiFr"}],"has_accepted_license":"1","date_updated":"2026-04-16T12:20:41Z","degree_awarded":"PhD","month":"10","ddc":["575"],"date_published":"2021-10-06T00:00:00Z","status":"public","citation":{"ieee":"L. Li, “Rapid cell growth regulation in Arabidopsis,” Institute of Science and Technology Austria, 2021.","ista":"Li L. 2021. Rapid cell growth regulation in Arabidopsis. Institute of Science and Technology Austria.","short":"L. Li, Rapid Cell Growth Regulation in Arabidopsis, Institute of Science and Technology Austria, 2021.","chicago":"Li, Lanxin. “Rapid Cell Growth Regulation in Arabidopsis.” Institute of Science and Technology Austria, 2021. <a href=\"https://doi.org/10.15479/at:ista:10083\">https://doi.org/10.15479/at:ista:10083</a>.","mla":"Li, Lanxin. <i>Rapid Cell Growth Regulation in Arabidopsis</i>. Institute of Science and Technology Austria, 2021, doi:<a href=\"https://doi.org/10.15479/at:ista:10083\">10.15479/at:ista:10083</a>.","apa":"Li, L. (2021). <i>Rapid cell growth regulation in Arabidopsis</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/at:ista:10083\">https://doi.org/10.15479/at:ista:10083</a>","ama":"Li L. Rapid cell growth regulation in Arabidopsis. 2021. doi:<a href=\"https://doi.org/10.15479/at:ista:10083\">10.15479/at:ista:10083</a>"},"doi":"10.15479/at:ista:10083","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","file":[{"date_updated":"2022-12-20T23:30:03Z","creator":"cchlebak","checksum":"3b2f55b3b8ae05337a0dcc1cd8595b10","content_type":"application/pdf","file_size":8616142,"file_id":"10138","relation":"main_file","date_created":"2021-10-14T08:00:07Z","access_level":"open_access","embargo":"2022-10-14","file_name":"0._IST_Austria_Thesis_Lanxin_Li_1014_pdftron.pdf"},{"relation":"source_file","access_level":"closed","date_created":"2021-10-14T08:00:13Z","file_name":"0._IST_Austria_Thesis_Lanxin_Li_1014.docx","date_updated":"2022-12-20T23:30:03Z","checksum":"f23ed258ca894f6aabf58b0c128bf242","creator":"cchlebak","embargo_to":"open_access","file_size":15058499,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"10139"}],"article_processing_charge":"No","year":"2021","abstract":[{"lang":"eng","text":"Plant motions occur across a wide spectrum of timescales, ranging from seed dispersal through bursting (milliseconds) and stomatal opening (minutes) to long-term adaptation of gross architecture. Relatively fast motions include water-driven growth as exemplified by root cell expansion under abiotic/biotic stresses or during gravitropism. A showcase is a root growth inhibition in 30 seconds triggered by the phytohormone auxin. However, the cellular and molecular mechanisms are still largely unknown. This thesis covers the studies about this topic as follows. By taking advantage of microfluidics combined with live imaging, pharmaceutical tools, and transgenic lines, we examined the kinetics of and causal relationship among various auxininduced rapid cellular changes in root growth, apoplastic pH, cytosolic Ca2+, cortical microtubule (CMT) orientation, and vacuolar morphology. We revealed that CMT reorientation and vacuolar constriction are the consequence of growth itself instead of responding directly to auxin. In contrast, auxin induces apoplast alkalinization to rapidly inhibit root growth in 30 seconds. This auxin-triggered apoplast alkalinization results from rapid H+- influx that is contributed by Ca2+ inward channel CYCLIC NUCLEOTIDE-GATED CHANNEL 14 (CNGC14)-dependent Ca2+ signaling. To dissect which auxin signaling mediates the rapid apoplast alkalinization, we\r\ncombined microfluidics and genetic engineering to verify that TIR1/AFB receptors conduct a non-transcriptional regulation on Ca2+ and H+ -influx. This non-canonical pathway is mostly mediated by the cytosolic portion of TIR1/AFB. On the other hand, we uncovered, using biochemical and phospho-proteomic analysis, that auxin cell surface signaling component TRANSMEMBRANE KINASE 1 (TMK1) plays a negative role during auxin-trigger apoplast\r\nalkalinization and root growth inhibition through directly activating PM H+ -ATPases. Therefore, we discovered that PM H+ -ATPases counteract instead of mediate the auxintriggered rapid H+ -influx, and that TIR1/AFB and TMK1 regulate root growth antagonistically. This opposite effect of TIR1/AFB and TMK1 is consistent during auxin-induced hypocotyl elongation, leading us to explore the relation of two signaling pathways. Assisted with biochemistry and fluorescent imaging, we verified for the first time that TIR1/AFB and TMK1 can interact with each other. The ability of TIR1/AFB binding to membrane lipid provides a basis for the interaction of plasma membrane- and cytosol-localized proteins.\r\nBesides, transgenic analysis combined with genetic engineering and biochemistry showed that  vi\r\nthey do function in the same pathway. Particularly, auxin-induced TMK1 increase is TIR1/AFB dependent, suggesting TIR1/AFB regulation on TMK1. Conversely, TMK1 also regulates TIR1/AFB protein levels and thus auxin canonical signaling. To follow the study of rapid growth regulation, we analyzed another rapid growth regulator, signaling peptide RALF1. We showed that RALF1 also triggers a rapid and reversible growth inhibition caused by H + influx, highly resembling but not dependent on auxin. Besides, RALF1 promotes auxin biosynthesis by increasing expression of auxin biosynthesis enzyme YUCCAs and thus induces auxin signaling in ca. 1 hour, contributing to the sustained RALF1-triggered growth inhibition. These studies collectively contribute to understanding rapid regulation on plant cell\r\ngrowth, novel auxin signaling pathway as well as auxin-peptide crosstalk. "}],"oa_version":"Published Version","_id":"10083","publication_status":"published","title":"Rapid cell growth regulation in Arabidopsis","ec_funded":1,"oa":1,"supervisor":[{"first_name":"Jiří","last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří"}],"publisher":"Institute of Science and Technology Austria","project":[{"grant_number":"665385","call_identifier":"H2020","name":"International IST Doctoral Program","_id":"2564DBCA-B435-11E9-9278-68D0E5697425"},{"name":"A Case Study of Plant Growth Regulation: Molecular Mechanism of Auxin-mediated Rapid Growth Inhibition in Arabidopsis Root","_id":"26B4D67E-B435-11E9-9278-68D0E5697425","grant_number":"25351"}],"tmp":{"short":"CC BY-NC-ND (4.0)","image":"/images/cc_by_nc_nd.png","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode"},"alternative_title":["ISTA Thesis"],"publication_identifier":{"issn":["2663-337X"]}},{"page":"164","status":"public","month":"09","date_published":"2020-09-30T00:00:00Z","ddc":["580"],"acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"}],"degree_awarded":"PhD","date_updated":"2026-07-28T12:51:58Z","department":[{"_id":"JiFr"}],"has_accepted_license":"1","type":"dissertation","corr_author":"1","acknowledgement":"I also want to thank the China Scholarship Council for supporting my study during the year from 2015 to 2019. I also want to thank IST facilities – the Bioimaging facility, the media kitchen, the plant facility and all of the campus services, for their support.","OA_place":"publisher","related_material":{"record":[{"id":"7643","status":"public","relation":"part_of_dissertation"}]},"file_date_updated":"2021-10-01T13:33:02Z","date_created":"2020-09-30T14:50:51Z","day":"30","language":[{"iso":"eng"}],"author":[{"full_name":"Han, Huibin","first_name":"Huibin","last_name":"Han","id":"31435098-F248-11E8-B48F-1D18A9856A87"}],"article_processing_charge":"No","file":[{"file_name":"2020_Han_Thesis.docx","date_created":"2020-09-30T14:50:20Z","access_level":"closed","relation":"source_file","file_id":"8590","file_size":49198118,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","creator":"dernst","checksum":"c4bda1947d4c09c428ac9ce667b02327","date_updated":"2020-09-30T14:50:20Z"},{"file_name":"2020_Han_Thesis.pdf","relation":"main_file","date_created":"2020-09-30T14:49:59Z","access_level":"open_access","file_size":15513963,"content_type":"application/pdf","file_id":"8591","date_updated":"2021-10-01T13:33:02Z","creator":"dernst","checksum":"3f4f5d1718c2230adf30639ecaf8a00b"}],"doi":"10.15479/AT:ISTA:8589","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","citation":{"mla":"Han, Huibin. <i>Novel Insights into PIN Polarity Regulation during Arabidopsis Development</i>. Institute of Science and Technology Austria, 2020, doi:<a href=\"https://doi.org/10.15479/AT:ISTA:8589\">10.15479/AT:ISTA:8589</a>.","apa":"Han, H. (2020). <i>Novel insights into PIN polarity regulation during Arabidopsis development</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT:ISTA:8589\">https://doi.org/10.15479/AT:ISTA:8589</a>","ama":"Han H. Novel insights into PIN polarity regulation during Arabidopsis development. 2020. doi:<a href=\"https://doi.org/10.15479/AT:ISTA:8589\">10.15479/AT:ISTA:8589</a>","chicago":"Han, Huibin. “Novel Insights into PIN Polarity Regulation during Arabidopsis Development.” Institute of Science and Technology Austria, 2020. <a href=\"https://doi.org/10.15479/AT:ISTA:8589\">https://doi.org/10.15479/AT:ISTA:8589</a>.","short":"H. Han, Novel Insights into PIN Polarity Regulation during Arabidopsis Development, Institute of Science and Technology Austria, 2020.","ista":"Han H. 2020. Novel insights into PIN polarity regulation during Arabidopsis development. Institute of Science and Technology Austria.","ieee":"H. Han, “Novel insights into PIN polarity regulation during Arabidopsis development,” Institute of Science and Technology Austria, 2020."},"oa":1,"title":"Novel insights into PIN polarity regulation during Arabidopsis development","publication_status":"published","_id":"8589","oa_version":"Published Version","abstract":[{"text":"The plant hormone auxin plays indispensable roles in plant growth and development. An essential level of regulation in auxin action is the directional auxin transport within cells. The establishment of auxin gradient in plant tissue has been attributed to local auxin biosynthesis and directional intercellular auxin transport, which both are controlled by various environmental and developmental signals. It is well established that asymmetric auxin distribution in cells is achieved by polarly localized PIN-FORMED (PIN) auxin efflux transporters. Despite the initial insights into cellular mechanisms of PIN polarization obtained from the last decades, the molecular mechanism and specific regulators mediating PIN polarization remains elusive. In this thesis, we aim to find novel players in PIN subcellular polarity regulation during Arabidopsis development. We first characterize the physiological effect of piperonylic acid (PA) on Arabidopsis hypocotyl gravitropic bending and PIN polarization. Secondly, we reveal the importance of SCFTIR1/AFB auxin signaling pathway in shoot gravitropism bending termination. In addition, we also explore the role of myosin XI complex, and actin cytoskeleton in auxin feedback regulation on PIN polarity. In Chapter 1, we give an overview of the current knowledge about PIN-mediated auxin fluxes in various plant tropic responses. In Chapter 2, we study the physiological effect of PA on shoot gravitropic bending. Our results show that PA treatment inhibits auxin-mediated PIN3 repolarization by interfering with PINOID and PIN3 phosphorylation status, ultimately leading to hyperbending hypocotyls. In Chapter 3, we provide evidence to show that the SCFTIR1/AFB nuclear auxin signaling pathway is crucial and required for auxin-mediated PIN3 repolarization and shoot gravitropic bending termination. In Chapter 4, we perform a phosphoproteomics approach and identify the motor protein Myosin XI and its binding protein, the MadB2 family, as an essential regulator of PIN polarity for auxin-canalization related developmental processes. In Chapter 5, we demonstrate the vital role of actin cytoskeleton in auxin feedback on PIN polarity by regulating PIN subcellular trafficking. Overall, the data presented in this PhD thesis brings novel insights into the PIN polar localization regulation that resulted in the (re)establishment of the polar auxin flow and gradient in response to environmental stimuli during plant development.","lang":"eng"}],"year":"2020","publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"publisher":"Institute of Science and Technology Austria","supervisor":[{"first_name":"Jiří","last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří"}]},{"article_processing_charge":"No","file":[{"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_size":91279806,"file_id":"8919","date_updated":"2021-07-16T22:30:03Z","embargo_to":"open_access","creator":"jhajny","checksum":"210a9675af5e4c78b0b56d920ac82866","file_name":"Jakub Hajný IST Austria final_JH.docx","relation":"source_file","date_created":"2020-12-04T07:27:52Z","access_level":"closed"},{"embargo":"2021-12-07","file_name":"Jakub Hajný IST Austria final_JH-merged without Science.pdf","relation":"main_file","date_created":"2020-12-09T15:04:41Z","access_level":"open_access","file_id":"8933","content_type":"application/pdf","file_size":68707697,"creator":"jhajny","checksum":"1781385b4aa73eba89cc76c6172f71d2","date_updated":"2021-12-08T23:30:03Z"}],"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT:ISTA:8822","citation":{"chicago":"Hajny, Jakub. “Identification and Characterization of the Molecular Machinery of Auxin-Dependent Canalization during Vasculature Formation and Regeneration.” Institute of Science and Technology Austria, 2020. <a href=\"https://doi.org/10.15479/AT:ISTA:8822\">https://doi.org/10.15479/AT:ISTA:8822</a>.","mla":"Hajny, Jakub. <i>Identification and Characterization of the Molecular Machinery of Auxin-Dependent Canalization during Vasculature Formation and Regeneration</i>. Institute of Science and Technology Austria, 2020, doi:<a href=\"https://doi.org/10.15479/AT:ISTA:8822\">10.15479/AT:ISTA:8822</a>.","apa":"Hajny, J. (2020). <i>Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT:ISTA:8822\">https://doi.org/10.15479/AT:ISTA:8822</a>","ama":"Hajny J. Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration. 2020. doi:<a href=\"https://doi.org/10.15479/AT:ISTA:8822\">10.15479/AT:ISTA:8822</a>","ieee":"J. Hajny, “Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration,” Institute of Science and Technology Austria, 2020.","ista":"Hajny J. 2020. Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration. Institute of Science and Technology Austria.","short":"J. Hajny, Identification and Characterization of the Molecular Machinery of Auxin-Dependent Canalization during Vasculature Formation and Regeneration, Institute of Science and Technology Austria, 2020."},"page":"249","status":"public","ddc":["580"],"date_published":"2020-12-01T00:00:00Z","month":"12","degree_awarded":"PhD","date_updated":"2026-06-18T19:02:05Z","has_accepted_license":"1","department":[{"_id":"JiFr"}],"type":"dissertation","corr_author":"1","OA_place":"publisher","related_material":{"record":[{"status":"public","id":"449","relation":"part_of_dissertation"},{"id":"7500","status":"public","relation":"part_of_dissertation"},{"id":"7427","status":"public","relation":"part_of_dissertation"},{"id":"191","status":"public","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","id":"6260","status":"public"}]},"file_date_updated":"2021-12-08T23:30:03Z","date_created":"2020-12-01T12:38:18Z","language":[{"iso":"eng"}],"day":"01","author":[{"first_name":"Jakub","last_name":"Hajny","id":"4800CC20-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0003-2140-7195","full_name":"Hajny, Jakub"}],"publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"publisher":"Institute of Science and Technology Austria","supervisor":[{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"}],"oa":1,"title":"Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration","_id":"8822","publication_status":"published","oa_version":"Published Version","abstract":[{"text":"Self-organization is a hallmark of plant development manifested e.g. by intricate leaf vein patterns, flexible formation of vasculature during organogenesis or its regeneration following wounding. Spontaneously arising channels transporting the phytohormone auxin, created by coordinated polar localizations of PIN-FORMED 1 (PIN1) auxin exporter, provide positional cues for these as well as other plant patterning processes. To find regulators acting downstream of auxin and the TIR1/AFB auxin signaling pathway essential for PIN1 coordinated polarization during auxin canalization, we performed microarray experiments. Besides the known components of general PIN polarity maintenance, such as PID and PIP5K kinases, we identified and characterized a new regulator of auxin canalization, the transcription factor WRKY DNA-BINDING PROTEIN 23 (WRKY23).\r\nNext, we designed a subsequent microarray experiment to further uncover other molecular players, downstream of auxin-TIR1/AFB-WRKY23 involved in the regulation of auxin-mediated PIN repolarization. We identified a novel and crucial part of the molecular machinery underlying auxin canalization. The auxin-regulated malectin-type receptor-like kinase CAMEL and the associated leucine-rich repeat receptor-like kinase CANAR target and directly phosphorylate PIN auxin transporters. camel and canar mutants are impaired in PIN1 subcellular trafficking and auxin-mediated repolarization leading to defects in auxin transport, ultimately to leaf venation and vasculature regeneration defects. Our results describe the CAMEL-CANAR receptor complex, which is required for auxin feed-back on its own transport and thus for coordinated tissue polarization during auxin canalization.","lang":"eng"}],"year":"2020"},{"year":"2019","abstract":[{"text":"The development and growth of Arabidopsis thaliana is regulated by a combination of genetic programing and also by the environmental influences. An important role in these processes play the phytohormones and among them, auxin is crucial as it controls many important functions. It is transported through the whole plant body by creating local and temporal concentration maxima and minima, which have an impact on the cell status, tissue and organ identity. Auxin has the property to undergo a directional and finely regulated cell-to-cell transport, which is enabled by the transport proteins, localized on the plasma membrane. An important role in this process have the PIN auxin efflux proteins, which have an asymmetric/polar subcellular localization and determine the directionality of the auxin transport. During the last years, there were significant advances in understanding how the trafficking molecular machineries function, including studies on molecular interactions, function, subcellular localization and intracellular distribution. However, there is still a lack of detailed characterization on the steps of endocytosis, exocytosis, endocytic recycling and degradation. Due to this fact, I focused on the identification of novel trafficking factors and better characterization of the intracellular trafficking pathways. My PhD thesis consists of an introductory chapter, three experimental chapters, a chapter containing general discussion, conclusions and perspectives and also an appendix chapter with published collaborative papers.\r\nThe first chapter is separated in two different parts: I start by a general introduction to auxin biology and then I introduce the trafficking pathways in the model plant Arabidopsis thaliana. Then, I explain also the phosphorylation-signals for polar targeting and also the roles of the phytohormone strigolactone.\r\nThe second chapter includes the characterization of bar1/sacsin mutant, which was identified in a forward genetic screen for novel trafficking components in Arabidopsis thaliana, where by the implementation of an EMS-treated pPIN1::PIN1-GFP marker line and by using the established inhibitor of ARF-GEFs, Brefeldin A (BFA) as a tool to study trafficking processes, we identified a novel factor, which is mediating the adaptation of the plant cell to ARF-GEF inhibition. The mutation is in a previously uncharacterized gene, encoding a very big protein that we, based on its homologies, called SACSIN with domains suggesting roles as a molecular chaperon or as a component of the ubiquitin-proteasome system. Our physiology and imaging studies revealed that SACSIN is a crucial plant cell component of the adaptation to the ARF-GEF inhibition.\r\nThe third chapter includes six subchapters, where I focus on the role of the phytohormone strigolactone, which interferes with auxin feedback on PIN internalization. Strigolactone moderates the polar auxin transport by increasing the internalization of the PIN auxin efflux carriers, which reduces the canalization related growth responses. In addition, I also studied the role of phosphorylation in the strigolactone regulation of auxin feedback on PIN internalization. In this chapter I also present my results on the MAX2-dependence of strigolactone-mediated root growth inhibition and I also share my results on the auxin metabolomics profiling after application of GR24.\r\nIn the fourth chapter I studied the effect of two small molecules ES-9 and ES9-17, which were identified from a collection of small molecules with the property to impair the clathrin-mediated endocytosis.\r\nIn the fifth chapter, I discuss all my observations and experimental findings and suggest alternative hypothesis to interpret my results.\r\nIn the appendix there are three collaborative published projects. In the first, I participated in the characterization of the role of ES9 as a small molecule, which is inhibitor of clathrin- mediated endocytosis in different model organisms. In the second paper, I contributed to the characterization of another small molecule ES9-17, which is a non-protonophoric analog of ES9 and also impairs the clathrin-mediated endocytosis not only in plant cells, but also in mammalian HeLa cells. Last but not least, I also attach another paper, where I tried to establish the grafting method as a technique in our lab to study canalization related processes.","lang":"eng"}],"oa":1,"publication_status":"published","_id":"7172","title":"Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana","oa_version":"Published Version","publisher":"Institute of Science and Technology Austria","supervisor":[{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"}],"publication_identifier":{"eissn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"corr_author":"1","related_material":{"record":[{"relation":"part_of_dissertation","id":"449","status":"public"},{"status":"public","id":"6377","relation":"part_of_dissertation"},{"relation":"part_of_dissertation","id":"1346","status":"public"}]},"OA_place":"publisher","language":[{"iso":"eng"}],"date_created":"2019-12-11T21:24:39Z","day":"12","file_date_updated":"2020-07-14T12:47:51Z","author":[{"last_name":"Vasileva","id":"3407EB18-F248-11E8-B48F-1D18A9856A87","first_name":"Mina K","full_name":"Vasileva, Mina K"}],"status":"public","page":"192","acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"Bio"}],"date_published":"2019-12-12T00:00:00Z","ddc":["570"],"month":"12","date_updated":"2026-04-08T13:54:45Z","degree_awarded":"PhD","type":"dissertation","department":[{"_id":"JiFr"}],"has_accepted_license":"1","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT:ISTA:7172","citation":{"chicago":"Vasileva, Mina K. “Molecular Mechanisms of Endomembrane Trafficking in Arabidopsis Thaliana.” Institute of Science and Technology Austria, 2019. <a href=\"https://doi.org/10.15479/AT:ISTA:7172\">https://doi.org/10.15479/AT:ISTA:7172</a>.","mla":"Vasileva, Mina K. <i>Molecular Mechanisms of Endomembrane Trafficking in Arabidopsis Thaliana</i>. Institute of Science and Technology Austria, 2019, doi:<a href=\"https://doi.org/10.15479/AT:ISTA:7172\">10.15479/AT:ISTA:7172</a>.","apa":"Vasileva, M. K. (2019). <i>Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT:ISTA:7172\">https://doi.org/10.15479/AT:ISTA:7172</a>","ama":"Vasileva MK. Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana. 2019. doi:<a href=\"https://doi.org/10.15479/AT:ISTA:7172\">10.15479/AT:ISTA:7172</a>","ieee":"M. K. Vasileva, “Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana,” Institute of Science and Technology Austria, 2019.","short":"M.K. Vasileva, Molecular Mechanisms of Endomembrane Trafficking in Arabidopsis Thaliana, Institute of Science and Technology Austria, 2019.","ista":"Vasileva MK. 2019. Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana. Institute of Science and Technology Austria."},"file":[{"relation":"source_file","date_created":"2019-12-12T09:32:36Z","access_level":"closed","file_name":"Thesis_Mina_final_upload_7.docx","creator":"mvasilev","checksum":"ef981c1a3b1d9da0edcbedcff4970d37","date_updated":"2020-07-14T12:47:51Z","file_id":"7175","file_size":20454014,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document"},{"file_id":"7176","file_size":11565025,"content_type":"application/pdf","checksum":"3882c4585e46c9cfb486e4225cad54ab","creator":"mvasilev","date_updated":"2020-07-14T12:47:51Z","file_name":"Thesis_Mina_final_upload_7.pdf","access_level":"open_access","date_created":"2019-12-12T09:33:10Z","relation":"main_file"}],"article_processing_charge":"No"},{"year":"2019","abstract":[{"lang":"eng","text":"Clathrin-Mediated Endocytosis (CME) is an aspect of cellular trafficking that is constantly regulated for mediating developmental and physiological responses. The main aim of my thesis is to decipher the basic mechanisms of CME and post-endocytic trafficking in the whole multicellular organ systems of Arabidopsis. The first chapter of my thesis describes the search for new components involved in CME. Tandem affinity purification was conducted using CLC and its interacting partners were identified. Amongst the identified proteins were the Auxilin-likes1 and 2 (Axl1/2), putative uncoating factors, for which we made a full functional analysis. Over-expression of Axl1/2 causes extreme modifications in the dynamics of the machinery proteins and inhibition of endocytosis altogether. However the loss of function of the axl1/2 did not present any cellular or physiological phenotype, meaning Auxilin-likes do not form the major uncoating machinery. The second chapter of my thesis describes the establishment/utilisation of techniques to capture the dynamicity and the complexity of CME and post-endocytic trafficking. We have studied the development of endocytic pits at the PM – specifically, the mode of membrane remodeling during pit development and the role of actin in it, given plant cells possess high turgor pressure. Utilizing the improved z-resolution of TIRF and VAEM techniques, we captured the time-lapse of the endocytic events at the plasma membrane; and using particle detection software, we quantitatively analysed all the endocytic trajectories in an unbiased way to obtain the endocytic rate of the system. This together with the direct analysis of cargo internalisation from the PM provided an estimate on the endocytic potential of the cell. We also developed a methodology for ultrastructural analysis of different populations of Clathrin-Coated Structures (CCSs) in both PM and endomembranes in unroofed protoplasts. Structural analysis, together with the intensity profile of CCSs at the PM show that the mode of CCP development at the PM follows ‘Constant curvature model’; meaning that clathrin polymerisation energy is a major contributing factor of membrane remodeling. In addition, other analyses clearly show that actin is not required for membrane remodeling during invagination or any other step of CCP development, despite the prevalent high turgor pressure. However, actin is essential in orchestrating the post-endocytic trafficking of CCVs facilitating the EE formation. We also observed that the uncoating process post-endocytosis is not immediate; an alternative mechanism of uncoating – Sequential multi-step process – functions in the cell. Finally we also looked at one of the important physiological stimuli modulating the process – hormone, auxin. auxin has been known to influence CME before. We have made a detailed study on the concentration-time based effect of auxin on the machinery proteins, CCP development, and the specificity of cargoes endocytosed. To this end, we saw no general effect of auxin on CME at earlier time points. However, very low concentration of IAA, such as 50nM, accelerates endocytosis of specifically PIN2 through CME. Such a tight regulatory control with high specificity to PIN2 could be essential in modulating its polarity. "}],"oa_version":"Published Version","publication_status":"published","_id":"6269","title":"Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants ","oa":1,"supervisor":[{"last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"}],"publisher":"Institute of Science and Technology Austria","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png","short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"alternative_title":["ISTA Thesis"],"publication_identifier":{"issn":["2663-337X"]},"author":[{"full_name":"Narasimhan, Madhumitha","orcid":"0000-0002-8600-0671","id":"44BF24D0-F248-11E8-B48F-1D18A9856A87","last_name":"Narasimhan","first_name":"Madhumitha"}],"day":"04","language":[{"iso":"eng"}],"date_created":"2019-04-09T14:37:06Z","file_date_updated":"2021-02-11T23:30:15Z","related_material":{"record":[{"status":"public","id":"412","relation":"part_of_dissertation"}]},"OA_place":"publisher","corr_author":"1","type":"dissertation","department":[{"_id":"JiFr"}],"has_accepted_license":"1","date_updated":"2026-04-08T14:00:24Z","degree_awarded":"PhD","date_published":"2019-02-04T00:00:00Z","month":"02","acknowledged_ssus":[{"_id":"Bio"},{"_id":"EM-Fac"}],"ddc":["575"],"status":"public","page":"138","citation":{"chicago":"Narasimhan, Madhumitha. “Clathrin-Mediated Endocytosis, Post-Endocytic Trafficking and Their Regulatory Controls in Plants .” Institute of Science and Technology Austria, 2019. <a href=\"https://doi.org/10.15479/at:ista:th1075\">https://doi.org/10.15479/at:ista:th1075</a>.","ama":"Narasimhan M. Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants . 2019. doi:<a href=\"https://doi.org/10.15479/at:ista:th1075\">10.15479/at:ista:th1075</a>","apa":"Narasimhan, M. (2019). <i>Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants </i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/at:ista:th1075\">https://doi.org/10.15479/at:ista:th1075</a>","mla":"Narasimhan, Madhumitha. <i>Clathrin-Mediated Endocytosis, Post-Endocytic Trafficking and Their Regulatory Controls in Plants </i>. Institute of Science and Technology Austria, 2019, doi:<a href=\"https://doi.org/10.15479/at:ista:th1075\">10.15479/at:ista:th1075</a>.","ieee":"M. Narasimhan, “Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants ,” Institute of Science and Technology Austria, 2019.","ista":"Narasimhan M. 2019. Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants . Institute of Science and Technology Austria.","short":"M. Narasimhan, Clathrin-Mediated Endocytosis, Post-Endocytic Trafficking and Their Regulatory Controls in Plants , Institute of Science and Technology Austria, 2019."},"doi":"10.15479/at:ista:th1075","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","file":[{"relation":"main_file","date_created":"2019-04-09T14:35:18Z","access_level":"open_access","file_name":"Supplementary_movie_1.avi","embargo":"2020-02-11","date_updated":"2021-02-11T23:30:15Z","creator":"dernst","checksum":"c958f27dd752712886e7e2638b847a3c","file_size":5402078,"content_type":"video/x-msvideo","file_id":"6270"},{"relation":"main_file","access_level":"open_access","date_created":"2019-04-09T14:35:18Z","file_name":"3.7_supplementary_movie_10.avi","embargo":"2020-02-11","date_updated":"2021-02-11T23:30:15Z","checksum":"8786fdc29c62987c0aad3c866a4d3691","creator":"dernst","file_size":5927736,"content_type":"video/x-msvideo","file_id":"6271"},{"content_type":"video/x-msvideo","file_size":9570210,"file_id":"6272","date_updated":"2021-02-11T23:30:15Z","creator":"dernst","checksum":"25f784c5159d6f4d966b2f9b371ebaf6","file_name":"3.7_supplementary_movie_9.avi","embargo":"2020-02-11","relation":"main_file","date_created":"2019-04-09T14:35:18Z","access_level":"open_access"},{"embargo":"2020-02-11","file_name":"3.7_supplementary_movie_8.avi","relation":"main_file","date_created":"2019-04-09T14:35:18Z","access_level":"open_access","file_id":"6273","content_type":"video/x-msvideo","file_size":2827360,"checksum":"917069272a7a08d1f38224d5e12765d6","creator":"dernst","date_updated":"2021-02-11T23:30:15Z"},{"file_name":"3.7_supplementary_movie_7.avi","embargo":"2020-02-11","relation":"main_file","access_level":"open_access","date_created":"2019-04-09T14:35:18Z","content_type":"video/x-msvideo","file_size":5771410,"file_id":"6274","date_updated":"2021-02-11T23:30:15Z","checksum":"81e74f5ca0ad70050504f18192236dc0","creator":"dernst"},{"file_name":"3.7_supplementary_movie_6.avi","embargo":"2020-02-11","relation":"main_file","date_created":"2019-04-09T14:35:18Z","access_level":"open_access","file_id":"6275","file_size":1113486,"content_type":"video/x-msvideo","checksum":"47eb37b27a2930252713924307ea8c6f","creator":"dernst","date_updated":"2021-02-11T23:30:15Z"},{"date_updated":"2021-02-11T23:30:15Z","creator":"dernst","checksum":"f68f66721041ce84e331959c9a5779c3","content_type":"video/x-msvideo","file_size":1057232,"file_id":"6276","relation":"main_file","date_created":"2019-04-09T14:35:18Z","access_level":"open_access","file_name":"3.7_supplementary_movie_5.avi","embargo":"2020-02-11"},{"date_updated":"2021-02-11T23:30:15Z","checksum":"67c01cefab51b363c5e214fe4cd671f3","creator":"dernst","file_size":127472916,"content_type":"video/x-msvideo","file_id":"6277","date_created":"2019-04-09T14:35:23Z","access_level":"open_access","relation":"main_file","embargo":"2020-02-11","file_name":"3.7_supplementary_movie_3.avi"},{"file_id":"6278","file_size":3181238,"content_type":"video/x-msvideo","creator":"dernst","checksum":"e5a397edbee05b8821e2b19b3c1a9260","date_updated":"2021-02-11T23:30:15Z","file_name":"3.7_supplementary_movie_4.avi","embargo":"2020-02-11","relation":"main_file","access_level":"open_access","date_created":"2019-04-09T14:35:19Z"},{"file_id":"6279","content_type":"video/x-msvideo","file_size":5970952,"creator":"dernst","checksum":"32d92b2a9277f956fdb0b42351d07c0b","date_updated":"2021-02-11T23:30:15Z","embargo":"2020-02-11","file_name":"3.7_supplementary_movie_2.avi","date_created":"2019-04-09T14:35:19Z","access_level":"open_access","relation":"main_file"},{"date_updated":"2021-02-11T23:30:15Z","creator":"dernst","checksum":"efe7001f5d9a8c61e631e12d5f324ade","content_type":"video/x-msvideo","file_size":39835236,"file_id":"6280","date_created":"2019-04-09T14:35:21Z","access_level":"open_access","relation":"main_file","file_name":"3.7_Supplementary_movie_1.avi","embargo":"2020-02-11"},{"file_name":"2.5_Suppl_Movie_4_AP2A1_TagRFP.avi","embargo":"2020-02-11","relation":"main_file","date_created":"2019-04-09T14:35:21Z","access_level":"open_access","file_id":"6281","content_type":"video/x-msvideo","file_size":3696740,"checksum":"eeb0a5603c6449c5f34eacd5ff0b3a16","creator":"dernst","date_updated":"2021-02-11T23:30:15Z"},{"file_name":"2.5_Suppl_Movie_3_TPLATE_GFP.avi","embargo":"2020-02-11","relation":"main_file","access_level":"open_access","date_created":"2019-04-09T14:35:21Z","file_id":"6282","file_size":6741232,"content_type":"video/x-msvideo","checksum":"8e7c00ef6223bf0e177deb168338af13","creator":"dernst","date_updated":"2021-02-11T23:30:15Z"},{"access_level":"open_access","date_created":"2019-04-09T14:35:22Z","relation":"main_file","embargo":"2020-02-11","file_name":"2.5_Suppl_Movie_2_CLC_GFP.avi","date_updated":"2021-02-11T23:30:15Z","checksum":"3636006a7cb709a7543d6581e359b28d","creator":"dernst","file_size":2445946,"content_type":"video/x-msvideo","file_id":"6283"},{"creator":"dernst","checksum":"39ca5519a6e9a38356e7b3704004fea7","date_updated":"2021-02-11T23:30:15Z","file_id":"6284","content_type":"video/x-msvideo","file_size":58594,"relation":"main_file","date_created":"2019-04-09T14:35:22Z","access_level"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Thesis"],"publication_identifier":{"issn":["2663-337X"]},"publisher":"Institute of Science and Technology Austria","supervisor":[{"id":"4159519E-F248-11E8-B48F-1D18A9856A87","last_name":"Friml","first_name":"Jiří","full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596"}],"oa":1,"oa_version":"Published Version","publication_status":"published","_id":"938","title":"Investigations into cell polarity and trafficking in the plant model Arabidopsis thaliana ","pubrep_id":"842","year":"2017","abstract":[{"lang":"eng","text":"The thesis encompasses several topics of plant cell biology which were studied in the model plant Arabidopsis thaliana. Chapter 1 concerns the plant hormone auxin and its polar transport through cells and tissues. The highly controlled, directional transport of auxin is facilitated by plasma membrane-localized transporters. Transporters from the PIN family direct auxin transport due to their polarized localizations at cell membranes. Substantial effort has been put into research on cellular trafficking of PIN proteins, which is thought to underlie their polar distribution. I participated in a forward genetic screen aimed at identifying novel regulators of PIN polarity. The screen yielded several genes which may be involved in PIN polarity regulation or participate in polar auxin transport by other means. Chapter 2 focuses on the endomembrane system, with particular attention to clathrin-mediated endocytosis. The project started with identification of several proteins that interact with clathrin light chains. Among them, I focused on two putative homologues of auxilin, which in non-plant systems is an endocytotic factor known for uncoating clathrin-coated vesicles in the final step of endocytosis. The body of my work consisted of an in-depth characterization of transgenic A. thaliana lines overexpressing these putative auxilins in an inducible manner. Overexpression of these proteins leads to an inhibition of endocytosis, as documented by imaging of cargoes and clathrin-related endocytic machinery. An extension of this work is an investigation into a concept of homeostatic regulation acting between distinct transport processes in the endomembrane system. With auxilin overexpressing lines, where endocytosis is blocked specifically, I made observations on the mutual relationship between two opposite trafficking processes of secretion and endocytosis. In Chapter 3, I analyze cortical microtubule arrays and their relationship to auxin signaling and polarized growth in elongating cells. In plants, microtubules are organized into arrays just below the plasma membrane, and it is thought that their function is to guide membrane-docked cellulose synthase complexes. These, in turn, influence cell wall structure and cell shape by directed deposition of cellulose fibres. In elongating cells, cortical microtubule arrays are able to reorient in relation to long cell axis, and these reorientations have been linked to cell growth and to signaling of growth-regulating factors such as auxin or light. In this chapter, I am addressing the causal relationship between microtubule array reorientation, growth, and auxin signaling. I arrive at a model where array reorientation is not guided by auxin directly, but instead is only controlled by growth, which, in turn, is regulated by auxin."}],"file":[{"file_size":46903863,"content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document","file_id":"6215","date_updated":"2020-07-14T12:48:15Z","creator":"dernst","checksum":"193425764d9aaaed3ac57062a867b315","file_name":"2017_Adamowski-Thesis_Source.docx","access_level":"closed","date_created":"2019-04-05T09:03:20Z","relation":"source_file"},{"file_id":"6216","file_size":8698888,"content_type":"application/pdf","creator":"dernst","checksum":"df5ab01be81f821e1b958596a1ec8d21","date_updated":"2020-07-14T12:48:15Z","file_name":"2017_Adamowski-Thesis.pdf","access_level":"open_access","date_created":"2019-04-05T09:03:19Z","relation":"main_file"}],"article_processing_charge":"No","publist_id":"6483","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","doi":"10.15479/AT:ISTA:th_842","citation":{"ieee":"M. Adamowski, “Investigations into cell polarity and trafficking in the plant model Arabidopsis thaliana ,” Institute of Science and Technology Austria, 2017.","short":"M. Adamowski, Investigations into Cell Polarity and Trafficking in the Plant Model Arabidopsis Thaliana , Institute of Science and Technology Austria, 2017.","ista":"Adamowski M. 2017. Investigations into cell polarity and trafficking in the plant model Arabidopsis thaliana . Institute of Science and Technology Austria.","chicago":"Adamowski, Maciek. “Investigations into Cell Polarity and Trafficking in the Plant Model Arabidopsis Thaliana .” Institute of Science and Technology Austria, 2017. <a href=\"https://doi.org/10.15479/AT:ISTA:th_842\">https://doi.org/10.15479/AT:ISTA:th_842</a>.","mla":"Adamowski, Maciek. <i>Investigations into Cell Polarity and Trafficking in the Plant Model Arabidopsis Thaliana </i>. Institute of Science and Technology Austria, 2017, doi:<a href=\"https://doi.org/10.15479/AT:ISTA:th_842\">10.15479/AT:ISTA:th_842</a>.","ama":"Adamowski M. Investigations into cell polarity and trafficking in the plant model Arabidopsis thaliana . 2017. doi:<a href=\"https://doi.org/10.15479/AT:ISTA:th_842\">10.15479/AT:ISTA:th_842</a>","apa":"Adamowski, M. (2017). <i>Investigations into cell polarity and trafficking in the plant model Arabidopsis thaliana </i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT:ISTA:th_842\">https://doi.org/10.15479/AT:ISTA:th_842</a>"},"date_published":"2017-06-02T00:00:00Z","month":"06","ddc":["581","583","580"],"status":"public","page":"117","type":"dissertation","department":[{"_id":"JiFr"}],"has_accepted_license":"1","date_updated":"2026-04-08T14:20:45Z","degree_awarded":"PhD","related_material":{"record":[{"relation":"part_of_dissertation","status":"public","id":"1591"}]},"OA_place":"publisher","corr_author":"1","author":[{"orcid":"0000-0001-6463-5257","full_name":"Adamowski, Maciek","first_name":"Maciek","last_name":"Adamowski","id":"45F536D2-F248-11E8-B48F-1D18A9856A87"}],"day":"02","date_created":"2018-12-11T11:49:18Z","language":[{"iso":"eng"}],"file_date_updated":"2020-07-14T12:48:15Z"},{"date_updated":"2026-07-29T12:01:27Z","degree_awarded":"PhD","type":"dissertation","has_accepted_license":"1","department":[{"_id":"JiFr"},{"_id":"GradSch"}],"status":"public","page":"131","date_published":"2017-01-12T00:00:00Z","ddc":["580"],"month":"01","day":"12","date_created":"2018-12-11T11:50:17Z","language":[{"iso":"eng"}],"file_date_updated":"2021-02-22T11:52:56Z","author":[{"last_name":"Prat","id":"3DA3BFEE-F248-11E8-B48F-1D18A9856A87","first_name":"Tomas","full_name":"Prat, Tomas"}],"corr_author":"1","doi_confirm":"1","acknowledgement":"I would like to first acknowledge my supervisor Jiří Friml for support, kind advice and patience. It was a pleasure to be a part of your lab, Jiří. I will remember the atmosphere present in auxin lab at VIB in Ghent and at IST in Klosterneuburg forever. I would like to thank all past and present lab members for the friendship and friendly and scientific environment in the groups. It was so nice to cooperate with you, guys. There was always someone who helped me with experiments, troubleshoot issues coming from our work etc. At this place, I would like to thank especially to Gergo Molnár. I’m happy (and lucky) that I have met him; he naturally became my tutor and guide through my PhD. From no one else during my entire professional career, I’ve learned that much.","related_material":{"record":[{"relation":"part_of_dissertation","status":"public","id":"449"}]},"OA_place":"publisher","file":[{"file_name":"IST_Austria_Thesis_Tomáš_Prát.pdf","date_created":"2019-04-05T08:45:14Z","access_level":"closed","relation":"main_file","file_id":"6209","file_size":10285946,"content_type":"application/pdf","creator":"dernst","checksum":"d192c7c6c5ea32c8432437286dc4909e","date_updated":"2019-04-05T08:45:14Z"},{"file_name":"2017_Thesis_Prat.pdf","success":1,"relation":"main_file","date_created":"2021-02-22T11:52:56Z","access_level":"open_access","file_id":"9185","file_size":9802991,"content_type":"application/pdf","checksum":"bab18b52cf98145926042d8ed99fdb3b","creator":"dernst","date_updated":"2021-02-22T11:52:56Z"}],"article_processing_charge":"No","citation":{"chicago":"Prat, Tomas. “Identification of Novel Regulators of PIN Polarity and Development of Novel Auxin Sensor.” Institute of Science and Technology Austria, 2017.","mla":"Prat, Tomas. <i>Identification of Novel Regulators of PIN Polarity and Development of Novel Auxin Sensor</i>. Institute of Science and Technology Austria, 2017.","apa":"Prat, T. (2017). <i>Identification of novel regulators of PIN polarity and development of novel auxin sensor</i>. Institute of Science and Technology Austria.","ama":"Prat T. Identification of novel regulators of PIN polarity and development of novel auxin sensor. 2017.","ieee":"T. Prat, “Identification of novel regulators of PIN polarity and development of novel auxin sensor,” Institute of Science and Technology Austria, 2017.","short":"T. Prat, Identification of Novel Regulators of PIN Polarity and Development of Novel Auxin Sensor, Institute of Science and Technology Austria, 2017.","ista":"Prat T. 2017. Identification of novel regulators of PIN polarity and development of novel auxin sensor. Institute of Science and Technology Austria."},"user_id":"8b945eb4-e2f2-11eb-945a-df72226e66a9","publist_id":"6233","publication_status":"published","_id":"1127","title":"Identification of novel regulators of PIN polarity and development of novel auxin sensor","oa_version":"Published Version","oa":1,"year":"2017","abstract":[{"text":"Plant hormone auxin and its transport between cells belong to the most important\r\nmechanisms controlling plant development. Auxin itself could change localization of PINs and\r\nthereby control direction of its own flow. We performed an expression profiling experiment\r\nin Arabidopsis roots to identify potential regulators of PIN polarity which are transcriptionally\r\nregulated by auxin signalling. We identified several novel regulators and performed a detailed\r\ncharacterization of the transcription factor WRKY23 (At2g47260) and its role in auxin\r\nfeedback on PIN polarity. Gain-of-function and dominant-negative mutants revealed that\r\nWRKY23 plays a crucial role in mediating the auxin effect on PIN polarity. In concordance,\r\ntypical polar auxin transport processes such as gravitropism and leaf vascular pattern\r\nformation were disturbed by interfering with WRKY23 function.\r\nIn order to identify direct targets of WRKY23, we performed consequential expression\r\nprofiling experiments using a WRKY23 inducible gain-of-function line and dominant-negative\r\nWRKY23 line that is defunct in PIN re-arrangement. Among several genes mostly related to\r\nthe groups of cell wall and defense process regulators, we identified LYSINE-HISTIDINE\r\nTRANSPORTER 1 (LHT1; At5g40780), a small amino acid permease gene from the amino\r\nacid/auxin permease family (AAAP), we present its detailed characterisation in auxin feedback\r\non PIN repolarization, identified its transcriptional regulation, we propose a potential\r\nmechanism of its action. Moreover, we identified also a member of receptor-like protein\r\nkinase LRR-RLK (LEUCINE-RICH REPEAT TRANSMEMBRANE PROTEIN KINASE PROTEIN 1;\r\nLRRK1; At1g05700), which also affects auxin-dependent PIN re-arrangement. We described\r\nits transcriptional behaviour, subcellular localization. Based on global expression data, we\r\ntried to identify ligand responsible for mechanism of signalling and suggest signalling partner\r\nand interactors. Additionally, we described role of novel phytohormone group, strigolactone,\r\nin auxin-dependent PIN re-arrangement, that could be a fundament for future studies in this\r\nfield.\r\nOur results provide first insights into an auxin transcriptional network targeting PIN\r\nlocalization and thus regulating plant development. We highlighted WRKY23 transcriptional\r\nnetwork and characterised its mediatory role in plant development. We identified direct\r\neffectors of this network, LHT1 and LRRK1, and describe their roles in PIN re-arrangement and\r\nPIN-dependent auxin transport processes.","lang":"eng"}],"publication_identifier":{"issn":["2663-337X"]},"alternative_title":["ISTA Thesis"],"supervisor":[{"first_name":"Jiří","last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8302-7596","full_name":"Friml, Jiří"}],"publisher":"Institute of Science and Technology Austria"},{"citation":{"chicago":"Marhavá, Petra. “Molecular Mechanisms of Patterning and Subcellular Trafficking in Arabidopsis Thaliana.” Institute of Science and Technology Austria, 2014.","mla":"Marhavá, Petra. <i>Molecular Mechanisms of Patterning and Subcellular Trafficking in Arabidopsis Thaliana</i>. Institute of Science and Technology Austria, 2014.","apa":"Marhavá, P. (2014). <i>Molecular mechanisms of patterning and subcellular trafficking in Arabidopsis thaliana</i>. Institute of Science and Technology Austria.","ama":"Marhavá P. Molecular mechanisms of patterning and subcellular trafficking in Arabidopsis thaliana. 2014.","ieee":"P. Marhavá, “Molecular mechanisms of patterning and subcellular trafficking in Arabidopsis thaliana,” Institute of Science and Technology Austria, 2014.","short":"P. Marhavá, Molecular Mechanisms of Patterning and Subcellular Trafficking in Arabidopsis Thaliana, Institute of Science and Technology Austria, 2014.","ista":"Marhavá P. 2014. Molecular mechanisms of patterning and subcellular trafficking in Arabidopsis thaliana. Institute of Science and Technology Austria."},"supervisor":[{"full_name":"Friml, Jiří","orcid":"0000-0002-8302-7596","last_name":"Friml","id":"4159519E-F248-11E8-B48F-1D18A9856A87","first_name":"Jiří"}],"publist_id":"5805","publisher":"Institute of Science and Technology Austria","user_id":"8b945eb4-e2f2-11eb-945a-df72226e66a9","alternative_title":["ISTA Thesis"],"article_processing_charge":"No","publication_identifier":{"issn":["2663-337X"]},"author":[{"full_name":"Marhavá, Petra","last_name":"Marhavá","id":"44E59624-F248-11E8-B48F-1D18A9856A87","first_name":"Petra"}],"date_created":"2018-12-11T11:51:49Z","day":"01","language":[{"iso":"eng"}],"OA_place":"publisher","year":"2014","doi_confirm":"1","corr_author":"1","abstract":[{"lang":"eng","text":"Phosphatidylinositol (Ptdlns) is a structural phospholipid that can be phosphorylated into various lipid signaling molecules, designated polyphosphoinositides (PPIs). The reversible phosphorylation of PPIs on the 3, 4, or 5 position of inositol is performed by a set of organelle-specific kinases and phosphatases, and the characteristic head groups make these molecules ideal for regulating biological processes in time and space. In yeast and mammals, Ptdlns3P and Ptdlns(3,5)P2 play crucial roles in trafficking toward the lytic compartments, whereas the role in plants is not yet fully understood. Here we identified the role of a land plant-specific subgroup of PPI phosphatases, the suppressor of actin 2 (SAC2) to SAC5, during vauolar trafficking and morphogenesis in Arabidopsis thaliana. SAC2-SAC5 localize to the tonoplast along with Ptdlns3P, the presumable product of their activity. in SAC gain- and loss-of-function mutants, the levels of Ptdlns monophosphates and bisphosphates were changed, with opposite effects on the morphology of storage and lytic vacuoles, and the trafficking toward the vacuoles was defective. Moreover, multiple sac knockout mutants had an increased number of smaller storage and lytic vacuoles, whereas extralarge vacuoles were observed in the overexpression lines, correlating with various growth and developmental defects. The fragmented vacuolar phenotype of sac mutants could be mimicked by treating wild-type seedlings with Ptdlns(3,5)P2, corroborating that this PPI is important for vacuole morphology. Taken together, these results provide evidence that PPIs, together with their metabolic enzymes SAC2-SAC5, are crucial for vacuolar trafficking and for vacuolar morphology and function in plants."}],"type":"dissertation","department":[{"_id":"JiFr"},{"_id":"GradSch"}],"oa_version":"None","date_updated":"2026-07-29T10:06:44Z","_id":"1402","publication_status":"published","degree_awarded":"PhD","title":"Molecular mechanisms of patterning and subcellular trafficking in Arabidopsis thaliana","month":"12","date_published":"2014-12-01T00:00:00Z","status":"public","page":"90"}]