[{"OA_type":"gold","supplementarymaterial":"yes","acknowledgement":"We thank A. Salmazo for assistance with Pol II purification. We thank staff at the Vienna BioCenter Core Facilities (VBCF) Proteomics facility for immunoprecipitation-mass spectrometry analysis, and J.A. Stopp for assistance with IP-MS data visualization. This research was further supported by the Scientific Service Units (SSUs) of ISTA through resources provided by the Lab Support Facility (LSF), Electron Microscopy Facility (EMF), Scientific Computing (SciComp), and the Preclinical Facility (PCF). F.H. was funded by the Endowed Professorship of the Lower Austria Research Funding Agency (GFF NÖ) and by the Austrian Research Promotion Agency (FFG) through the COIN Establishment Grant n.o. 45624401.","type":"journal_article","oa_version":"Published Version","das_tickbox":"1","biorxivid":1,"researchdata_availability":"yes","OA_place":"publisher","DOAJ_listed":"1","department":[{"_id":"CaBe"}],"external_id":{"biorxivid":["10.64898/2025.12.10.692585"]},"publication_identifier":{"eissn":["2041-1723"]},"ddc":["570"],"oa":1,"article_type":"original","language":[{"iso":"eng"}],"status":"public","PlanS_conform":"1","date_updated":"2026-07-16T11:29:31Z","_id":"22333","scopus_import":"1","author":[{"id":"36062FEC-F248-11E8-B48F-1D18A9856A87","last_name":"Hlavata","full_name":"Hlavata, Annamaria","first_name":"Annamaria"},{"last_name":"Neuditschko","full_name":"Neuditschko, Benjamin","first_name":"Benjamin"},{"last_name":"Schellhaas","full_name":"Schellhaas, Ulla","first_name":"Ulla"},{"last_name":"Plaschka","full_name":"Plaschka, Clemens","first_name":"Clemens"},{"last_name":"Herzog","full_name":"Herzog, Franz","first_name":"Franz"},{"id":"2CB9DFE2-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0003-0893-7036","full_name":"Bernecky, Carrie A","last_name":"Bernecky","first_name":"Carrie A"}],"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","publication_status":"epub_ahead","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"main_file_link":[{"url":"https://doi.org/10.1038/s41467-026-75416-8","open_access":"1"}],"date_created":"2026-07-14T07:27:59Z","article_processing_charge":"Yes","dataavailabilitystatement":"The\r\nc ryo EM maps generated in this study were deposited to the EM Data Bank under the\r\naccession codes: EMD 55583 [https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55583\r\n(Pol II Gdown1 RPAP2 composite map), EMD 55578\r\n[https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55578 Pol II Gdown1 RPAP2 Pol II core\r\nmap EMD 55579 [https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55579 Pol II\r\nGdown1 RPAP2 Pol II stalk map EMD 55580\r\n[https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55580 Pol II Gdown1 RPAP2 RPAP2\r\nmap EMD 55581 [https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55 581 Pol II\r\nGdown1 RPAP2 Gdown1 N terminus map EMD 55582\r\n[https://www.ebi.ac.uk/pdbe/entry/emdb/EMD 55582 Pol II Gdown1 RPAP2 Gdown1\r\nC terminus map and EMD 55585 [https://www.ebi.ac.uk/pdbe/entry/emdb/EMD\r\n55585 RPAP2 GPN1 GPN3 map Model coordi nates were deposited to the PDBe under\r\nthe accession codes: 9T5H [http://doi.org/10.2210/pdb 9T5H / (Pol II Gdown1\r\nRPAP2 complex structure) and 9T5J [http://doi.org/10.2210/pdb 9T5H / (GPN1\r\nGPN3 RPAP2 structure). Immunoprecipitation mass spectrometry and crosslinking mass\r\nspectrometry proteomics data have been deposited to the ProteomeXchange Consortium\r\nvia the PRIDE partner repository with the dataset identifiers PXD071638\r\n[http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD 071638 and\r\nP XD070852\r\n[http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD 070852\r\nAlphaFold3 structure predictions have been deposited to the Zenodo repository\r\nhttps://doi.org/10.5281/zenodo.20687910 P reviously published model coordinates\r\nwere utilized and are available at the PDB under the accession codes 8QEP\r\n[http://doi.org/10.2210/pdb 8QEP / 9BZ 0 [http://doi.org/10.2210/pdb 9BZ 0 /\r\nand 7B7U [http://doi.org/10.2210/pdb 7B7U / Source Data are provided with this\r\npaper.","abstract":[{"text":"RNA polymerase II (Pol II) must be assembled in the cytoplasm before it enters the nucleus, where it transcribes protein-coding genes. Although transcription by Pol II is intensively studied, how this central multi-subunit enzyme is made and the role of dedicated assembly factors remains unclear. Here, we report the integrative structural analysis of a native human Pol II from the cytoplasm captured near the end of biogenesis. The complex contains Gdown1 and three biogenesis factors – RPAP2 and the critical small GTPases GPN1 and GPN3. Cryo-EM analysis of the complex reveals how Gdown1 and RPAP2 associate with Pol II and prevent the premature association of transcription factors. Further biochemical and cryo-EM analysis reveals how RPAP2 tethers GPN1–GPN3 to the complex and how the assembly of the RPAP2–GPN1–GPN3 complex is controlled by GTP hydrolysis. The combined results uncover a network of interactions that chaperone cytoplasmic Pol II to prevent aberrant interactions, reveal a molecular switch regulating biogenesis factor association, and suggest a general mechanism for the action of GPN-loop GTPase family of enzymes.","lang":"eng"}],"citation":{"ieee":"A. Hlavata, B. Neuditschko, U. Schellhaas, C. Plaschka, F. Herzog, and C. Bernecky, “Structure of cytoplasmic RNA polymerase II,” <i>Nature Communications</i>. Springer Nature, 2026.","chicago":"Hlavata, Annamaria, Benjamin Neuditschko, Ulla Schellhaas, Clemens Plaschka, Franz Herzog, and Carrie Bernecky. “Structure of Cytoplasmic RNA Polymerase II.” <i>Nature Communications</i>. Springer Nature, 2026. <a href=\"https://doi.org/10.1038/s41467-026-75416-8\">https://doi.org/10.1038/s41467-026-75416-8</a>.","ista":"Hlavata A, Neuditschko B, Schellhaas U, Plaschka C, Herzog F, Bernecky C. 2026. Structure of cytoplasmic RNA polymerase II. Nature Communications.","apa":"Hlavata, A., Neuditschko, B., Schellhaas, U., Plaschka, C., Herzog, F., &#38; Bernecky, C. (2026). Structure of cytoplasmic RNA polymerase II. <i>Nature Communications</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41467-026-75416-8\">https://doi.org/10.1038/s41467-026-75416-8</a>","short":"A. Hlavata, B. Neuditschko, U. Schellhaas, C. Plaschka, F. Herzog, C. Bernecky, Nature Communications (2026).","ama":"Hlavata A, Neuditschko B, Schellhaas U, Plaschka C, Herzog F, Bernecky C. Structure of cytoplasmic RNA polymerase II. <i>Nature Communications</i>. 2026. doi:<a href=\"https://doi.org/10.1038/s41467-026-75416-8\">10.1038/s41467-026-75416-8</a>","mla":"Hlavata, Annamaria, et al. “Structure of Cytoplasmic RNA Polymerase II.” <i>Nature Communications</i>, Springer Nature, 2026, doi:<a href=\"https://doi.org/10.1038/s41467-026-75416-8\">10.1038/s41467-026-75416-8</a>."},"corr_author":"1","acknowledged_ssus":[{"_id":"LifeSc"},{"_id":"EM-Fac"},{"_id":"ScienComp"},{"_id":"PreCl"}],"title":"Structure of cytoplasmic RNA polymerase II","publication":"Nature Communications","quality_controlled":"1","license":"https://creativecommons.org/licenses/by/4.0/","date_published":"2026-07-13T00:00:00Z","day":"13","year":"2026","month":"07","publisher":"Springer Nature","doi":"10.1038/s41467-026-75416-8","has_accepted_license":"1"}]