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TitleThe human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans.
Journal, issue, pagesLife Sci Alliance, Vol. 5, Issue 11, Year 2022
Publish dateSep 1, 2022
AuthorsJulia L Daiß / Michael Pilsl / Kristina Straub / Andrea Bleckmann / Mona Höcherl / Florian B Heiss / Guillermo Abascal-Palacios / Ewan P Ramsay / Katarina Tlučková / Jean-Clement Mars / Torben Fürtges / Astrid Bruckmann / Till Rudack / Carrie Bernecky / Valérie Lamour / Konstantin Panov / Alessandro Vannini / Tom Moss / Christoph Engel /
PubMed AbstractTranscription 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 ...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.
External linksLife Sci Alliance / PubMed:36271492 / PubMed Central
MethodsEM (single particle)
Resolution4.09 Å
Structure data

15135

8a43

EMDB-15135, PDB-8a43:
Human RNA polymerase I
Method: EM (single particle) / Resolution: 4.09 Å

Source
  • homo sapiens (human)
KeywordsTRANSCRIPTION / RNA polymerase I / human rDNA transcription

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