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

Julia 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

doi:10.26508/lsa.202201568

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

Julia 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^*

^*Corresponding author for this work

School of Biological Sciences

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English
Article number	e202201568
Journal	Life Science Alliance
Volume	5
Issue number	11
DOIs	https://doi.org/10.26508/lsa.202201568
Publication status	Published - 01 Sep 2022

Keywords

Animals
RNA Polymerase I - genetics - metabolism
DNA
Saccharomyces cerevisiae - metabolism
RNA Precursors
Transcription Factors - metabolism
Humans

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.26508/lsa.202201568Licence: CC BY

The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans
Copyright 2022 the authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited
Final published version, 3.04 MBLicence: CC BY

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Daiß, J. L., Pilsl, M., Straub, K., Bleckmann, A., Höcherl, M., Heiss, F. B., Abascal-Palacios, G., Ramsay, E. P., Tlučková, 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]. https://doi.org/10.26508/lsa.202201568

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

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.",

keywords = "Animals, RNA Polymerase I - genetics - metabolism, DNA, Saccharomyces cerevisiae - metabolism, RNA Precursors, Transcription Factors - metabolism, Humans",

author = "Dai{\ss}, {Julia L} and Michael Pilsl and Kristina Straub and Andrea Bleckmann and Mona H{\"o}cherl and Heiss, {Florian B} and Guillermo Abascal-Palacios and Ramsay, {Ewan P} and Katarina Tlu{\v c}kov{\'a} and Jean-Clement Mars and Torben F{\"u}rtges and Astrid Bruckmann and Till Rudack and Carrie Bernecky and Val{\'e}rie Lamour and Konstantin Panov and Alessandro Vannini and Tom Moss and Christoph Engel",

year = "2022",

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day = "1",

doi = "10.26508/lsa.202201568",

language = "English",

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Daiß, JL, Pilsl, M, Straub, K, Bleckmann, A, Höcherl, M, Heiss, FB, Abascal-Palacios, G, Ramsay, EP, Tlučková, 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, vol. 5, no. 11, e202201568. https://doi.org/10.26508/lsa.202201568

TY - JOUR

T1 - The human RNA polymerase I structure reveals an HMG-like docking domain specific to metazoans

AU - Daiß, Julia L

AU - Pilsl, Michael

AU - Straub, Kristina

AU - Bleckmann, Andrea

AU - Höcherl, Mona

AU - Heiss, Florian B

AU - Abascal-Palacios, Guillermo

AU - Ramsay, Ewan P

AU - Tlučková, Katarina

AU - Mars, Jean-Clement

AU - Fürtges, Torben

AU - Bruckmann, Astrid

AU - Rudack, Till

AU - Bernecky, Carrie

AU - Lamour, Valérie

AU - Panov, Konstantin

AU - Vannini, Alessandro

AU - Moss, Tom

AU - Engel, Christoph

PY - 2022/9/1

Y1 - 2022/9/1

N2 - 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.

AB - 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.

KW - Animals

KW - RNA Polymerase I - genetics - metabolism

KW - DNA

KW - Saccharomyces cerevisiae - metabolism

KW - RNA Precursors

KW - Transcription Factors - metabolism

KW - Humans

U2 - 10.26508/lsa.202201568

DO - 10.26508/lsa.202201568

M3 - Article

C2 - 36271492

VL - 5

JO - Life Science Alliance

JF - Life Science Alliance

SN - 2575-1077

IS - 11

M1 - e202201568

ER -

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

Abstract

Keywords

UN SDGs

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