[{"publisher":"Life Science Alliance","_id":"12051","scopus_import":"1","title":"The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans","status":"public","abstract":[{"text":"Transcription of the ribosomal RNA precursor by RNA polymerase (Pol) I is a major determinant of cellular growth, and dysregulation is observed in many cancer types. Here, we present the purification of human Pol I from cells carrying a genomic GFP fusion on the largest subunit allowing the structural and functional analysis of the enzyme across species. In contrast to yeast, human Pol I carries a single-subunit stalk, and in vitro transcription indicates a reduced proofreading activity. Determination of the human Pol I cryo-EM reconstruction in a close-to-native state rationalizes the effects of disease-associated mutations and uncovers an additional domain that is built into the sequence of Pol I subunit RPA1. This “dock II” domain resembles a truncated HMG box incapable of DNA binding which may serve as a downstream transcription factor–binding platform in metazoans. Biochemical analysis, in situ modelling, and ChIP data indicate that Topoisomerase 2a can be recruited to Pol I via the domain and cooperates with the HMG box domain–containing factor UBF. These adaptations of the metazoan Pol I transcription system may allow efficient release of positive DNA supercoils accumulating downstream of the transcription bubble.","lang":"eng"}],"doi":"10.26508/lsa.202201568","acknowledgement":"The authors especially thank Philip Gunkel for his contribution. We thank all\r\npast and present members of the Engel lab, Achim Griesenbeck, Colyn Crane-\r\nRobinson, Christophe Lotz, Marlene Vayssieres, Klaus Grasser, Herbert Tschochner, and Philipp Milkereit for help and discussion; Gerhard Lehmann and Nobert Eichner for IT support; Joost Zomerdijk for UBF-constructs, Volker Cordes for the Hela P2 cell line; Remco Sprangers for shared cell culture; Dina Grohmann and the Archaea Center for fermentation; and Thomas\r\nDresselhaus for access to fluorescence microscopes. This work was in part supported by the Emmy-Noether Programm (DFG grant no. EN 1204/1-1 to C Engel) of the German Research Council and Collaborative Research Center 960 (TP-A8 to C Engel).","type":"journal_article","file_date_updated":"2022-09-08T06:41:14Z","publication_identifier":{"issn":["2575-1077"]},"publication_status":"published","volume":5,"date_updated":"2024-10-21T06:01:48Z","article_number":"e202201568","oa":1,"file":[{"relation":"main_file","file_id":"12062","creator":"dernst","checksum":"4201d876a3e5e8b65e319d03300014ad","success":1,"access_level":"open_access","content_type":"application/pdf","date_updated":"2022-09-08T06:41:14Z","file_name":"2022_LifeScienceAlliance_Daiss.pdf","date_created":"2022-09-08T06:41:14Z","file_size":3183129}],"department":[{"_id":"CaBe"}],"oa_version":"Published Version","issue":"11","article_processing_charge":"No","day":"01","isi":1,"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","author":[{"last_name":"Daiß","first_name":"Julia L","full_name":"Daiß, Julia L"},{"first_name":"Michael","full_name":"Pilsl, Michael","last_name":"Pilsl"},{"last_name":"Straub","full_name":"Straub, Kristina","first_name":"Kristina"},{"first_name":"Andrea","full_name":"Bleckmann, Andrea","last_name":"Bleckmann"},{"full_name":"Höcherl, Mona","first_name":"Mona","last_name":"Höcherl"},{"last_name":"Heiss","full_name":"Heiss, Florian B","first_name":"Florian B"},{"last_name":"Abascal-Palacios","first_name":"Guillermo","full_name":"Abascal-Palacios, Guillermo"},{"last_name":"Ramsay","first_name":"Ewan P","full_name":"Ramsay, Ewan P"},{"last_name":"Tluckova","id":"4AC7D980-F248-11E8-B48F-1D18A9856A87","full_name":"Tluckova, Katarina","first_name":"Katarina"},{"last_name":"Mars","first_name":"Jean-Clement","full_name":"Mars, Jean-Clement"},{"last_name":"Fürtges","full_name":"Fürtges, Torben","first_name":"Torben"},{"last_name":"Bruckmann","first_name":"Astrid","full_name":"Bruckmann, Astrid"},{"last_name":"Rudack","full_name":"Rudack, Till","first_name":"Till"},{"id":"2CB9DFE2-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0003-0893-7036","last_name":"Bernecky","first_name":"Carrie A","full_name":"Bernecky, Carrie A"},{"last_name":"Lamour","first_name":"Valérie","full_name":"Lamour, Valérie"},{"last_name":"Panov","full_name":"Panov, Konstantin","first_name":"Konstantin"},{"first_name":"Alessandro","full_name":"Vannini, Alessandro","last_name":"Vannini"},{"first_name":"Tom","full_name":"Moss, Tom","last_name":"Moss"},{"last_name":"Engel","first_name":"Christoph","full_name":"Engel, Christoph"}],"tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png"},"keyword":["Health","Toxicology and Mutagenesis","Plant Science","Biochemistry","Genetics and Molecular Biology (miscellaneous)","Ecology"],"license":"https://creativecommons.org/licenses/by/4.0/","month":"09","year":"2022","publication":"Life Science Alliance","quality_controlled":"1","language":[{"iso":"eng"}],"external_id":{"isi":["000972702600001"]},"article_type":"original","intvolume":"         5","citation":{"ieee":"J. L. Daiß <i>et al.</i>, “The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans,” <i>Life Science Alliance</i>, vol. 5, no. 11. Life Science Alliance, 2022.","ama":"Daiß JL, Pilsl M, Straub K, et al. The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans. <i>Life Science Alliance</i>. 2022;5(11). doi:<a href=\"https://doi.org/10.26508/lsa.202201568\">10.26508/lsa.202201568</a>","ista":"Daiß JL, Pilsl M, Straub K, Bleckmann A, Höcherl M, Heiss FB, Abascal-Palacios G, Ramsay EP, Tluckova K, Mars J-C, Fürtges T, Bruckmann A, Rudack T, Bernecky C, Lamour V, Panov K, Vannini A, Moss T, Engel C. 2022. The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans. Life Science Alliance. 5(11), e202201568.","mla":"Daiß, Julia L., et al. “The Human RNA Polymerase I Structure Reveals an HMG-like Docking Domain Specific to Metazoans.” <i>Life Science Alliance</i>, vol. 5, no. 11, e202201568, Life Science Alliance, 2022, doi:<a href=\"https://doi.org/10.26508/lsa.202201568\">10.26508/lsa.202201568</a>.","apa":"Daiß, J. L., Pilsl, M., Straub, K., Bleckmann, A., Höcherl, M., Heiss, F. B., … Engel, C. (2022). The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans. <i>Life Science Alliance</i>. Life Science Alliance. <a href=\"https://doi.org/10.26508/lsa.202201568\">https://doi.org/10.26508/lsa.202201568</a>","chicago":"Daiß, Julia L, Michael Pilsl, Kristina Straub, Andrea Bleckmann, Mona Höcherl, Florian B Heiss, Guillermo Abascal-Palacios, et al. “The Human RNA Polymerase I Structure Reveals an HMG-like Docking Domain Specific to Metazoans.” <i>Life Science Alliance</i>. Life Science Alliance, 2022. <a href=\"https://doi.org/10.26508/lsa.202201568\">https://doi.org/10.26508/lsa.202201568</a>.","short":"J.L. Daiß, M. Pilsl, K. Straub, A. Bleckmann, M. Höcherl, F.B. Heiss, G. Abascal-Palacios, E.P. Ramsay, K. Tluckova, J.-C. Mars, T. Fürtges, A. Bruckmann, T. Rudack, C. Bernecky, V. Lamour, K. Panov, A. Vannini, T. Moss, C. Engel, Life Science Alliance 5 (2022)."},"has_accepted_license":"1","date_created":"2022-09-06T18:45:23Z","ddc":["570"],"date_published":"2022-09-01T00:00:00Z"},{"doi":"10.26508/lsa.201900623","abstract":[{"lang":"eng","text":"Nucleoporin 93 (Nup93) expression inversely correlates with the survival of triple-negative breast cancer patients. However, our knowledge of Nup93 function in breast cancer besides its role as structural component of the nuclear pore complex is not understood. Combination of functional assays and genetic analyses suggested that chromatin interaction of Nup93 partially modulates the expression of genes associated with actin cytoskeleton remodeling and epithelial to mesenchymal transition, resulting in impaired invasion of triple-negative, claudin-low breast cancer cells. Nup93 depletion induced stress fiber formation associated with reduced cell migration/proliferation and impaired expression of mesenchymal-like genes. Silencing LIMCH1, a gene responsible for actin cytoskeleton remodeling and up-regulated upon Nup93 depletion, partially restored the invasive phenotype of cancer cells. Loss of Nup93 led to significant defects in tumor establishment/propagation in vivo, whereas patient samples revealed that high Nup93 and low LIMCH1 expression correlate with late tumor stage. Our approach identified Nup93 as contributor of triple-negative, claudin-low breast cancer cell invasion and paves the way to study the role of nuclear envelope proteins during breast cancer tumorigenesis."}],"extern":"1","pmid":1,"volume":3,"date_updated":"2024-10-14T11:18:41Z","publication_status":"published","file_date_updated":"2022-04-08T07:33:01Z","publication_identifier":{"issn":["2575-1077"]},"type":"journal_article","scopus_import":"1","title":"Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling","publisher":"Life Science Alliance","_id":"11058","status":"public","quality_controlled":"1","language":[{"iso":"eng"}],"publication":"Life Science Alliance","year":"2020","citation":{"mla":"Bersini, Simone, et al. “Nup93 Regulates Breast Tumor Growth by Modulating Cell Proliferation and Actin Cytoskeleton Remodeling.” <i>Life Science Alliance</i>, vol. 3, no. 1, e201900623, Life Science Alliance, 2020, doi:<a href=\"https://doi.org/10.26508/lsa.201900623\">10.26508/lsa.201900623</a>.","apa":"Bersini, S., Lytle, N. K., Schulte, R., Huang, L., Wahl, G. M., &#38; Hetzer, M. (2020). Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling. <i>Life Science Alliance</i>. Life Science Alliance. <a href=\"https://doi.org/10.26508/lsa.201900623\">https://doi.org/10.26508/lsa.201900623</a>","chicago":"Bersini, Simone, Nikki K Lytle, Roberta Schulte, Ling Huang, Geoffrey M Wahl, and Martin Hetzer. “Nup93 Regulates Breast Tumor Growth by Modulating Cell Proliferation and Actin Cytoskeleton Remodeling.” <i>Life Science Alliance</i>. Life Science Alliance, 2020. <a href=\"https://doi.org/10.26508/lsa.201900623\">https://doi.org/10.26508/lsa.201900623</a>.","short":"S. Bersini, N.K. Lytle, R. Schulte, L. Huang, G.M. Wahl, M. Hetzer, Life Science Alliance 3 (2020).","ieee":"S. Bersini, N. K. Lytle, R. Schulte, L. Huang, G. M. Wahl, and M. Hetzer, “Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling,” <i>Life Science Alliance</i>, vol. 3, no. 1. Life Science Alliance, 2020.","ama":"Bersini S, Lytle NK, Schulte R, Huang L, Wahl GM, Hetzer M. Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling. <i>Life Science Alliance</i>. 2020;3(1). doi:<a href=\"https://doi.org/10.26508/lsa.201900623\">10.26508/lsa.201900623</a>","ista":"Bersini S, Lytle NK, Schulte R, Huang L, Wahl GM, Hetzer M. 2020. Nup93 regulates breast tumor growth by modulating cell proliferation and actin cytoskeleton remodeling. Life Science Alliance. 3(1), e201900623."},"date_created":"2022-04-07T07:44:18Z","has_accepted_license":"1","ddc":["570"],"date_published":"2020-01-01T00:00:00Z","article_type":"original","intvolume":"         3","external_id":{"pmid":["31959624"]},"oa_version":"Published Version","file":[{"creator":"dernst","checksum":"3bf33e7e93bef7823287807206b69b38","file_id":"11137","relation":"main_file","file_size":2653960,"date_updated":"2022-04-08T07:33:01Z","file_name":"2020_LifeScienceAlliance_Bersini.pdf","date_created":"2022-04-08T07:33:01Z","access_level":"open_access","success":1,"content_type":"application/pdf"}],"article_number":"e201900623","oa":1,"keyword":["Health","Toxicology and Mutagenesis","Plant Science","Biochemistry","Genetics and Molecular Biology (miscellaneous)","Ecology"],"month":"01","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png"},"author":[{"first_name":"Simone","full_name":"Bersini, Simone","last_name":"Bersini"},{"last_name":"Lytle","full_name":"Lytle, Nikki K","first_name":"Nikki K"},{"full_name":"Schulte, Roberta","first_name":"Roberta","last_name":"Schulte"},{"full_name":"Huang, Ling","first_name":"Ling","last_name":"Huang"},{"full_name":"Wahl, Geoffrey M","first_name":"Geoffrey M","last_name":"Wahl"},{"first_name":"Martin W","full_name":"HETZER, Martin W","last_name":"HETZER","orcid":"0000-0002-2111-992X","id":"86c0d31b-b4eb-11ec-ac5a-eae7b2e135ed"}],"article_processing_charge":"No","day":"01","issue":"1"}]