TY - JOUR

T1 - Regulation of the RNAPII Pool Is Integral to the DNA Damage Response

AU - Tufegdžić Vidaković, Ana

AU - Mitter, Richard

AU - Kelly, Gavin P.

AU - Neumann, Michelle

AU - Harreman, Michelle

AU - Rodríguez-Martínez, Marta

AU - Herlihy, Anna

AU - Weems, Juston C.

AU - Boeing, Stefan

AU - Encheva, Vesela

AU - Gaul, Liam

AU - Milligan, Laura

AU - Tollervey, David

AU - Conaway, Ronald C.

AU - Conaway, Joan W.

AU - Snijders, Ambrosius P.

AU - Stewart, Aengus

AU - Svejstrup, Jesper Q.

N1 - Funding Information: This work was supported by the Francis Crick Institute (FCI receives its core funding from Cancer Research UK [FC001166], the UK Medical Research Council [FC001166], and the Wellcome Trust [FC001166]), the European Research Council (Agreement 693327 to J.Q.S.), and the Wellcome Trust (1099160 to D.T.). We thank FCI's Advanced Sequencing Facility, Cell Services, and the High Throughput Screening Facility for their help with this project. The Flp-In-compatible destination vector was kindly provided by Markus Landthaler. The RNAPII CTD S2 phosphorylation-specific antibody (3E10) was a kind gift from Dirk Eick. We thank Eric Forgy for adapting his Pages.jl package, Melvin Gonzalez and Yanfeng He for their input, members of the Svejstrup laboratory for discussions, and Peter Verrijzer and Barbara Dirac-Svejstrup for helpful comments on the manuscript. A.T.V. and J.Q.S. conceived the project. A.T.V. M.N. M.H. M.R.M. A.H. J.C.W. S.B. V.E. L.G. and L.M. performed experiments. R.M. and A.T.V. performed data analysis. G.P.K. developed RNAPII mathematical modeling. D.T. R.C.C. J.W.C. A.P.S. A.S. and J.Q.S. supervised the different aspects of the work. A.T.V. and J.Q.S. wrote the manuscript, with input from all authors. The authors declare no competing interests. Funding Information: This work was supported by the Francis Crick Institute (FCI receives its core funding from Cancer Research UK [ FC001166 ], the UK Medical Research Council [ FC001166 ], and the Wellcome Trust [ FC001166 ]), the European Research Council (Agreement 693327 to J.Q.S.), and the Wellcome Trust ( 1099160 to D.T.). We thank FCI’s Advanced Sequencing Facility, Cell Services, and the High Throughput Screening Facility for their help with this project. The Flp-In-compatible destination vector was kindly provided by Markus Landthaler. The RNAPII CTD S2 phosphorylation-specific antibody (3E10) was a kind gift from Dirk Eick. We thank Eric Forgy for adapting his Pages.jl package, Melvin Gonzalez and Yanfeng He for their input, members of the Svejstrup laboratory for discussions, and Peter Verrijzer and Barbara Dirac-Svejstrup for helpful comments on the manuscript. Publisher Copyright: © 2020 The Author(s)

PY - 2020/3/19

Y1 - 2020/3/19

N2 - In response to transcription-blocking DNA damage, cells orchestrate a multi-pronged reaction, involving transcription-coupled DNA repair, degradation of RNA polymerase II (RNAPII), and genome-wide transcription shutdown. Here, we provide insight into how these responses are connected by the finding that ubiquitylation of RNAPII itself, at a single lysine (RPB1 K1268), is the focal point for DNA-damage-response coordination. K1268 ubiquitylation affects DNA repair and signals RNAPII degradation, essential for surviving genotoxic insult. RNAPII degradation results in a shutdown of transcriptional initiation, in the absence of which cells display dramatic transcriptome alterations. Additionally, regulation of RNAPII stability is central to transcription recovery—persistent RNAPII depletion underlies the failure of this process in Cockayne syndrome B cells. These data expose regulation of global RNAPII levels as integral to the cellular DNA-damage response and open the intriguing possibility that RNAPII pool size generally affects cell-specific transcription programs in genome instability disorders and even normal cells.

AB - In response to transcription-blocking DNA damage, cells orchestrate a multi-pronged reaction, involving transcription-coupled DNA repair, degradation of RNA polymerase II (RNAPII), and genome-wide transcription shutdown. Here, we provide insight into how these responses are connected by the finding that ubiquitylation of RNAPII itself, at a single lysine (RPB1 K1268), is the focal point for DNA-damage-response coordination. K1268 ubiquitylation affects DNA repair and signals RNAPII degradation, essential for surviving genotoxic insult. RNAPII degradation results in a shutdown of transcriptional initiation, in the absence of which cells display dramatic transcriptome alterations. Additionally, regulation of RNAPII stability is central to transcription recovery—persistent RNAPII depletion underlies the failure of this process in Cockayne syndrome B cells. These data expose regulation of global RNAPII levels as integral to the cellular DNA-damage response and open the intriguing possibility that RNAPII pool size generally affects cell-specific transcription programs in genome instability disorders and even normal cells.

KW - DNA damage

KW - RNA polymerase II

KW - transcription

KW - ubiquitin

KW - ubiquitylation

KW - UV irradiation

U2 - 10.1016/j.cell.2020.02.009

DO - 10.1016/j.cell.2020.02.009

M3 - Journal article

C2 - 32142654

AN - SCOPUS:85081645943

VL - 180

SP - 1245-1261.e21

JO - Cell

JF - Cell

SN - 0092-8674

IS - 6

ER -