TY - JOUR
T1 - ELOF1 is a transcription-coupled DNA repair factor that directs RNA polymerase II ubiquitylation
AU - van der Weegen, Yana
AU - de Lint, Klaas
AU - van den Heuvel, Diana
AU - Nakazawa, Yuka
AU - Mevissen, Tycho E. T.
AU - van Schie, Janne J. M.
AU - San Martin Alonso, Marta
AU - Boer, Daphne E. C.
AU - González-Prieto, Román
AU - Narayanan, Ishwarya V.
AU - Klaassen, Noud H. M.
AU - Wondergem, Annelotte P.
AU - Roohollahi, Khashayar
AU - Dorsman, Josephine C.
AU - Hara, Yuichiro
AU - Vertegaal, Alfred C. O.
AU - de Lange, Job
AU - Walter, Johannes C.
AU - Noordermeer, Sylvie M.
AU - Ljungman, Mats
AU - Ogi, Tomoo
AU - Wolthuis, Rob M. F.
AU - Luijsterburg, Martijn S.
N1 - Funding Information: We acknowledge A. Kragten, J. Balk, J. Poell, K. Kato, M. Shimada, S. Kloet, M. Paulsen, M. Vukic, D. Warmerdam, A. Ramadhin and L. Daxinger for help during this project. We also thank J. Moffat, K. Chan and A. Tong for sharing the pLCKO-TKOv3 library before publication. We thank the Amsterdam UMC NGS sequencing facilities for support. We thank P. van Veelen and A. de Ru for MS equipment maintenance. This work was funded by a LUMC Research Fellowship, a NWO-ENW-M grant (OCENW.KLEIN.090) and a NWO-VIDI grant (ALW.016.161.320) to M.S.L., a Leiden University Fund (LUF) grant to D.v.d.H. (W18355-2-EM), a KWF/Alpe Young Investigator 10701 grant to J.d.L., a CCA proof-of-concept grant to K.d.L. and R.W., an Amsterdam UMC Innovation Grant (CRISPR Expertise Center, 2019) to R.W., UM1 HG009382 and R01 CA213214 NCI grants to M.L., a KWF Young Investigator grant 11367 to R.G.-P., and an ERC starting grant 310913 to A.C.O.V. J.C.W. was supported by NIH grant HL098316 and is a Howard Hughes Medical Institute (HHMI) Investigator and an American Cancer Society Research Professor. T.E.T.M. was supported by an EMBO Long-term fellowship (ALTF 1316-2016) and a HHMI fellowship of The Jane Coffin Childs Memorial Fund for Medical Research. Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2021/6/1
Y1 - 2021/6/1
N2 - Cells employ transcription-coupled repair (TCR) to eliminate transcription-blocking DNA lesions. DNA damage-induced binding of the TCR-specific repair factor CSB to RNA polymerase II (RNAPII) triggers RNAPII ubiquitylation of a single lysine (K1268) by the CRL4CSA ubiquitin ligase. How CRL4CSA is specifically directed towards K1268 is unknown. Here, we identify ELOF1 as the missing link that facilitates RNAPII ubiquitylation, a key signal for the assembly of downstream repair factors. This function requires its constitutive interaction with RNAPII close to K1268, revealing ELOF1 as a specificity factor that binds and positions CRL4CSA for optimal RNAPII ubiquitylation. Drug–genetic interaction screening also revealed a CSB-independent pathway in which ELOF1 prevents R-loops in active genes and protects cells against DNA replication stress. Our study offers key insights into the molecular mechanisms of TCR and provides a genetic framework of the interplay between transcriptional stress responses and DNA replication.
AB - Cells employ transcription-coupled repair (TCR) to eliminate transcription-blocking DNA lesions. DNA damage-induced binding of the TCR-specific repair factor CSB to RNA polymerase II (RNAPII) triggers RNAPII ubiquitylation of a single lysine (K1268) by the CRL4CSA ubiquitin ligase. How CRL4CSA is specifically directed towards K1268 is unknown. Here, we identify ELOF1 as the missing link that facilitates RNAPII ubiquitylation, a key signal for the assembly of downstream repair factors. This function requires its constitutive interaction with RNAPII close to K1268, revealing ELOF1 as a specificity factor that binds and positions CRL4CSA for optimal RNAPII ubiquitylation. Drug–genetic interaction screening also revealed a CSB-independent pathway in which ELOF1 prevents R-loops in active genes and protects cells against DNA replication stress. Our study offers key insights into the molecular mechanisms of TCR and provides a genetic framework of the interplay between transcriptional stress responses and DNA replication.
UR - http://www.scopus.com/inward/record.url?scp=85107491310&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/s41556-021-00688-9
DO - https://doi.org/10.1038/s41556-021-00688-9
M3 - Article
C2 - 34108663
SN - 1465-7392
VL - 23
SP - 595
EP - 607
JO - Nature cell biology
JF - Nature cell biology
IS - 6
ER -