The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans

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.

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Journal Article | Published | English
Author
Daiß, Julia L; Pilsl, Michael; Straub, Kristina; Bleckmann, Andrea; Höcherl, Mona; Heiss, Florian B; Abascal-Palacios, Guillermo; Ramsay, Ewan P; Tluckova, KatarinaISTA; Mars, Jean-Clement; Fürtges, Torben; Bruckmann, Astrid
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Department
Abstract
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.
Publishing Year
Date Published
2022-09-01
Journal Title
Life Science Alliance
Publisher
Life Science Alliance
Acknowledgement
The authors especially thank Philip Gunkel for his contribution. We thank all past and present members of the Engel lab, Achim Griesenbeck, Colyn Crane- Robinson, 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 Dresselhaus 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).
Volume
5
Issue
11
Article Number
e202201568
ISSN
IST-REx-ID

Cite this

Daiß JL, Pilsl M, Straub K, et al. The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans. Life Science Alliance. 2022;5(11). doi:10.26508/lsa.202201568
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. Life Science Alliance. Life Science Alliance. https://doi.org/10.26508/lsa.202201568
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.” Life Science Alliance. Life Science Alliance, 2022. https://doi.org/10.26508/lsa.202201568.
J. L. Daiß et al., “The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans,” Life Science Alliance, vol. 5, no. 11. Life Science Alliance, 2022.
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.
Daiß, Julia L., et al. “The Human RNA Polymerase I Structure Reveals an HMG-like Docking Domain Specific to Metazoans.” Life Science Alliance, vol. 5, no. 11, e202201568, Life Science Alliance, 2022, doi:10.26508/lsa.202201568.
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2022-09-08
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