Catalytic and noncatalytic functions of DNA polymerase κ in translesion DNA synthesis.


Journal

Nature structural & molecular biology
ISSN: 1545-9985
Titre abrégé: Nat Struct Mol Biol
Pays: United States
ID NLM: 101186374

Informations de publication

Date de publication:
19 Sep 2024
Historique:
received: 01 09 2023
accepted: 28 08 2024
medline: 20 9 2024
pubmed: 20 9 2024
entrez: 19 9 2024
Statut: aheadofprint

Résumé

Translesion DNA synthesis (TLS) is a cellular process that enables the bypass of DNA lesions encountered during DNA replication and is emerging as a primary target of chemotherapy. Among vertebrate DNA polymerases, polymerase κ (Polκ) has the distinctive ability to bypass minor groove DNA adducts in vitro. However, Polκ is also required for cells to overcome major groove DNA adducts but the basis of this requirement is unclear. Here, we combine CRISPR base-editor screening technology in human cells with TLS analysis of defined DNA lesions in Xenopus egg extracts to unravel the functions and regulations of Polκ during lesion bypass. Strikingly, we show that Polκ has two main functions during TLS, which are differentially regulated by Rev1 binding. On the one hand, Polκ is essential to replicate across a minor groove DNA lesion in a process that depends on PCNA ubiquitylation but is independent of Rev1. On the other hand, through its cooperative interaction with Rev1 and ubiquitylated PCNA, Polκ appears to stabilize the Rev1-Polζ extension complex on DNA to allow extension past major groove DNA lesions and abasic sites, in a process that is independent of Polκ's catalytic activity. Together, our work identifies catalytic and noncatalytic functions of Polκ in TLS and reveals important regulatory mechanisms underlying the unique domain architecture present at the C-terminal end of Y-family TLS polymerases.

Identifiants

pubmed: 39300172
doi: 10.1038/s41594-024-01395-3
pii: 10.1038/s41594-024-01395-3
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

© 2024. The Author(s).

Références

Ling, H., Boudsocq, F., Woodgate, R. & Yang, W. Crystal structure of a Y-family DNA polymerase in action: a mechanism for error-prone and lesion-bypass replication. Cell 107, 91–102 (2001).
pubmed: 11595188 doi: 10.1016/S0092-8674(01)00515-3
Goodman, M. F. & Woodgate, R. Translesion DNA polymerases. Cold Spring Harb. Perspect. Biol. 5, a010363 (2013).
pubmed: 23838442 pmcid: 3783050 doi: 10.1101/cshperspect.a010363
Cortez, D. Replication-coupled DNA repair. Mol. Cell 74, 866–876 (2019).
pubmed: 31173722 pmcid: 6557297 doi: 10.1016/j.molcel.2019.04.027
Saldivar, J. C., Cortez, D. & Cimprich, K. A. The essential kinase ATR: ensuring faithful duplication of a challenging genome. Nat. Rev. Mol. Cell Biol. 18, 622–636 (2017).
pubmed: 28811666 pmcid: 5796526 doi: 10.1038/nrm.2017.67
Friedberg, E. C., Wagner, R. & Radman, M. Specialized DNA polymerases, cellular survival, and the genesis of mutations. Science 296, 1627–1630 (2002).
pubmed: 12040171 doi: 10.1126/science.1070236
Hoege, C., Pfander, B., Moldovan, G.-L., Pyrowolakis, G. & Jentsch, S. RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO. Nature 419, 135–141 (2002).
pubmed: 12226657 doi: 10.1038/nature00991
Stelter, P. & Ulrich, H. D. Control of spontaneous and damage-induced mutagenesis by SUMO and ubiquitin conjugation. Nature 425, 188–191 (2003).
pubmed: 12968183 doi: 10.1038/nature01965
Watanabe, K. et al. Rad18 guides Polη to replication stalling sites through physical interaction and PCNA monoubiquitination. EMBO J. 23, 3886–3896 (2004).
pubmed: 15359278 pmcid: 522788 doi: 10.1038/sj.emboj.7600383
Sale, J. E., Lehmann, A. R. & Woodgate, R. Y-family DNA polymerases and their role in tolerance of cellular DNA damage. Nat. Rev. Mol. Cell Biol. 13, 141–152 (2012).
pubmed: 22358330 pmcid: 3630503 doi: 10.1038/nrm3289
Johnson, R. E., Prakash, S. & Prakash, L. Efficient bypass of a thymine–thymine dimer by yeast DNA polymerase, Polη. Science 283, 1001–1004 (1999).
pubmed: 9974380 doi: 10.1126/science.283.5404.1001
Masutani, C., Kusumoto, R., Iwai, S. & Hanaoka, F. Mechanisms of accurate translesion synthesis by human DNA polymerase η. EMBO J. 19, 3100–3109 (2000).
pubmed: 10856253 pmcid: 203367 doi: 10.1093/emboj/19.12.3100
McCulloch, S. D. et al. Preferential cis–syn thymine dimer bypass by DNA polymerase η occurs with biased fidelity. Nature 428, 97–100 (2004).
pubmed: 14999287 doi: 10.1038/nature02352
Johnson, R. E., Washington, M. T., Haracska, L., Prakash, S. & Prakash, L. Eukaryotic polymerases ι and ζ act sequentially to bypass DNA lesions. Nature 406, 1015–1019 (2000).
pubmed: 10984059 doi: 10.1038/35023030
Lee, Y.-S., Gregory, M. T. & Yang, W. Human Pol ζ purified with accessory subunits is active in translesion DNA synthesis and complements Pol η in cisplatin bypass. Proc. Natl Acad. Sci. USA 111, 2954–2959 (2014).
pubmed: 24449906 pmcid: 3939873 doi: 10.1073/pnas.1324001111
Gallina, I. et al. The ubiquitin ligase RFWD3 is required for translesion DNA synthesis. Mol. Cell 81, 442–458 (2021).
pubmed: 33321094 pmcid: 7864614 doi: 10.1016/j.molcel.2020.11.029
Gibbs, P. E. M., McDonald, J., Woodgate, R. & Lawrence, C. W. The relative roles in vivo of Saccharomyces cerevisiae Pol η, Pol ζ, Rev1 protein and Pol32 in the bypass and mutation induction of an abasic site, T–T (6–4) photoadduct and T–T cis–syn cyclobutane dimer. Genetics 169, 575–582 (2005).
pubmed: 15520252 pmcid: 1449107 doi: 10.1534/genetics.104.034611
Hicks, J. K. et al. Differential roles for DNA polymerases η, ζ, and REV1 in lesion bypass of intrastrand versus interstrand DNA cross-links. Mol. Cell. Biol. 30, 1217–1230 (2010).
pubmed: 20028736 doi: 10.1128/MCB.00993-09
Yoon, J.-H., Prakash, L. & Prakash, S. Error-free replicative bypass of (6–4) photoproducts by DNA polymerase ζ in mouse and human cells. Genes Dev. 24, 123–128 (2010).
pubmed: 20080950 pmcid: 2807347 doi: 10.1101/gad.1872810
Guo, C. et al. REV1 protein interacts with PCNA: significance of the REV1 BRCT domain in vitro and in vivo. Mol. Cell 23, 265–271 (2006).
pubmed: 16857592 doi: 10.1016/j.molcel.2006.05.038
Martin, S. K. & Wood, R. D. DNA polymerase ζ in DNA replication and repair. Nucleic Acids Res. 47, 8348–8361 (2019).
pubmed: 31410467 pmcid: 6895278 doi: 10.1093/nar/gkz705
Acharya, N., Johnson, R. E., Pagès, V., Prakash, L. & Prakash, S. Yeast Rev1 protein promotes complex formation of DNA polymerase ζ with Pol32 subunit of DNA polymerase δ. Proc. Natl Acad. Sci. USA 106, 9631–9636 (2009).
pubmed: 19487673 pmcid: 2701015 doi: 10.1073/pnas.0902175106
Budzowska, M., Graham, T. G. W., Sobeck, A., Waga, S. & Walter, J. C. Regulation of the Rev1–Pol ζ complex during bypass of a DNA interstrand cross-link. EMBO J. 34, 1971–1985 (2015).
pubmed: 26071591 pmcid: 4547899 doi: 10.15252/embj.201490878
Edmunds, C. E., Simpson, L. J. & Sale, J. E. PCNA ubiquitination and REV1 define temporally distinct mechanisms for controlling translesion synthesis in the avian cell line DT40. Mol. Cell 30, 519–529 (2008).
pubmed: 18498753 doi: 10.1016/j.molcel.2008.03.024
Gerlach, V. L., Feaver, W. J., Fischhaber, P. L. & Friedberg, E. C. Purification and characterization of Polκ, a DNA polymerase encoded by the human DINB1 gene. J. Biol. Chem. 276, 92–98 (2001).
pubmed: 11024016 doi: 10.1074/jbc.M004413200
Ogi, T., Kato, T. Jr, Kato, T. & Ohmori, H. Mutation enhancement by DINB1, a mammalian homologue of the Escherichia coli mutagenesis protein dinB. Genes Cells 4, 607–618 (1999).
pubmed: 10620008 doi: 10.1046/j.1365-2443.1999.00289.x
Choi, J.-Y., Angel, K. C. & Guengerich, F. P. Translesion synthesis across bulky N
pubmed: 16751196 doi: 10.1074/jbc.M602246200
Jarosz, D. F., Godoy, V. G., Delaney, J. C., Essigmann, J. M. & Walker, G. C. A single amino acid governs enhanced activity of DinB DNA polymerases on damaged templates. Nature 439, 225–228 (2006).
pubmed: 16407906 doi: 10.1038/nature04318
Jha, V., Bian, C., Xing, G. & Ling, H. Structure and mechanism of error-free replication past the major benzo[a]pyrene adduct by human DNA polymerase κ. Nucleic Acids Res. 44, 4957–4967 (2016).
pubmed: 27034468 pmcid: 4889944 doi: 10.1093/nar/gkw204
Jha, V. & Ling, H. 2.0 Å resolution crystal structure of human Polκ reveals a new catalytic function of N-clasp in DNA replication. Sci Rep. 8, 15125 (2018).
pubmed: 30310122 pmcid: 6181923 doi: 10.1038/s41598-018-33371-5
Ogi, T., Shinkai, Y., Tanaka, K. & Ohmori, H. Polκ protects mammalian cells against the lethal and mutagenic effects of benzo[a]pyrene. Proc. Natl Acad. Sci. USA 99, 15548–15553 (2002).
pubmed: 12432099 pmcid: 137754 doi: 10.1073/pnas.222377899
Malvezzi, S. et al. Mechanism of RNA polymerase II stalling by DNA alkylation. Proc. Natl Acad. Sci. USA 114, 12172–12177 (2017).
pubmed: 29087308 pmcid: 5699039 doi: 10.1073/pnas.1706592114
Takeiri, A. et al. In vivo evidence that DNA polymerase κ is responsible for error-free bypass across DNA cross-links induced by mitomycin C. DNA Repair 24, 113–121 (2014).
pubmed: 25303778 doi: 10.1016/j.dnarep.2014.09.002
Tanasova, M. & Sturla, S. J. Chemistry and biology of acylfulvenes: sesquiterpene-derived antitumor agents. Chem. Rev. 112, 3578–3610 (2012).
pubmed: 22482429 doi: 10.1021/cr2001367
Casimir, L., Zimmer, S., Racine-Brassard, F., Jacques, P.-É. & Maréchal, A. The mutational impact of illudin S on human cells. DNA Repair 122, 103433 (2023).
pubmed: 36566616 doi: 10.1016/j.dnarep.2022.103433
Olivieri, M. et al. A genetic map of the response to DNA damage in human cells. Cell 182, 481–496 (2020).
pubmed: 32649862 pmcid: 7384976 doi: 10.1016/j.cell.2020.05.040
Williams, H. L., Gottesman, M. E. & Gautier, J. Replication-independent repair of DNA interstrand crosslinks. Mol. Cell 47, 140–147 (2012).
pubmed: 22658724 pmcid: 3666337 doi: 10.1016/j.molcel.2012.05.001
Ogi, T. & Lehmann, A. R. The Y-family DNA polymerase κ (Pol κ) functions in mammalian nucleotide-excision repair. Nat. Cell Biol. 8, 640–642 (2006).
pubmed: 16738703 doi: 10.1038/ncb1417
Spanjaard, A. et al. Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine. Nucleic Acids Res. 50, 7420–7435 (2022).
pubmed: 35819193 pmcid: 9303390 doi: 10.1093/nar/gkac545
Johnson, R. E., Prakash, S. & Prakash, L. The human DINB1 gene encodes the DNA polymerase Polθ. Proc. Natl Acad. Sci. USA 97, 3838–3843 (2000).
pubmed: 10760255 pmcid: 18103 doi: 10.1073/pnas.97.8.3838
Kanemaru, Y. et al. Catalytic and non-catalytic roles of DNA polymerase κ in the protection of human cells against genotoxic stresses. Environ. Mol. Mutagen. 56, 650–662 (2015).
pubmed: 26031400 doi: 10.1002/em.21961
Ogi, T. et al. Three DNA polymerases, recruited by different mechanisms, carry out NER repair synthesis in human cells. Mol. Cell 37, 714–727 (2010).
pubmed: 20227374 doi: 10.1016/j.molcel.2010.02.009
Bétous, R. et al. DNA polymerase κ-dependent DNA synthesis at stalled replication forks is important for CHK1 activation. EMBO J. 32, 2172–2185 (2013).
pubmed: 23799366 pmcid: 3730229 doi: 10.1038/emboj.2013.148
Dall’Osto, M., Pierini, L., Valery, N., Hoffmann, J.-S. & Pillaire, M.-J. A catalytically independent function of human DNA polymerase κ controls the stability and abundance of checkpoint kinase 1. Mol. Cell. Biol. 41, e0009021 (2021).
pubmed: 34398682 doi: 10.1128/MCB.00090-21
Tonzi, P., Yin, Y., Lee, C. W. T., Rothenberg, E. & Huang, T. T. Translesion polymerase κ-dependent DNA synthesis underlies replication fork recovery. eLife 7, e41426 (2018).
pubmed: 30422114 pmcid: 6251625 doi: 10.7554/eLife.41426
Hishiki, A. et al. Structural basis for novel interactions between human translesion synthesis polymerases and proliferating cell nuclear antigen. J. Biol. Chem. 284, 10552–10560 (2009).
pubmed: 19208623 pmcid: 2667742 doi: 10.1074/jbc.M809745200
Vaisman, A. & Woodgate, R. Translesion DNA polymerases in eukaryotes: what makes them tick? Crit. Rev. Biochem. Mol. Biol. 52, 274–303 (2017).
pubmed: 28279077 pmcid: 5573590 doi: 10.1080/10409238.2017.1291576
Xie, W., Yang, X., Xu, M. & Jiang, T. Structural insights into the assembly of human translesion polymerase complexes. Protein Cell 3, 864–874 (2012).
pubmed: 23143872 pmcid: 4875464 doi: 10.1007/s13238-012-2102-x
Sangree, A. K. et al. Benchmarking of SpCas9 variants enables deeper base editor screens of BRCA1 and BCL2. Nat. Commun. 13, 1318 (2022).
pubmed: 35288574 pmcid: 8921519 doi: 10.1038/s41467-022-28884-7
Boudsocq, F. et al. Investigating the role of the little finger domain of Y-family DNA polymerases in low fidelity synthesis and translesion replication. J. Biol. Chem. 279, 32932–32940 (2004).
pubmed: 15155753 doi: 10.1074/jbc.M405249200
Walter, J., Sun, L. & Newport, J. Regulated chromosomal DNA replication in the absence of a nucleus. Mol. Cell 1, 519–529 (1998).
pubmed: 9660936 doi: 10.1016/S1097-2765(00)80052-0
Ogi, T., Kannouche, P. & Lehmann, A. R. Localisation of human Y-family DNA polymerase κ: relationship to PCNA foci. J. Cell Sci. 118, 129–136 (2005).
pubmed: 15601657 doi: 10.1242/jcs.01603
Guo, C., Tang, T.-S., Bienko, M., Dikic, I. & Friedberg, E. C. Requirements for the interaction of mouse Polκ with ubiquitin and its biological significance. J. Biol. Chem. 283, 4658–4664 (2008).
pubmed: 18162470 doi: 10.1074/jbc.M709275200
Ohashi, E. et al. Identification of a novel REV1-interacting motif necessary for DNA polymerase κ function. Genes Cells 14, 101–111 (2009).
pubmed: 19170759 pmcid: 3103050 doi: 10.1111/j.1365-2443.2008.01255.x
Masuda, Y. et al. Different types of interaction between PCNA and PIP boxes contribute to distinct cellular functions of Y-family DNA polymerases. Nucleic Acids Res. 43, 7898–7910 (2015).
pubmed: 26170230 pmcid: 4652755 doi: 10.1093/nar/gkv712
Lancey, C. et al. Cryo-EM structure of human Pol κ bound to DNA and mono-ubiquitylated PCNA. Nat. Commun. 12, 6095 (2021).
pubmed: 34667155 pmcid: 8526622 doi: 10.1038/s41467-021-26251-6
Larsen, N. B. et al. Replication-coupled DNA–protein crosslink repair by SPRTN and the proteasome in Xenopus egg extracts. Mol. Cell 73, 574–588 (2019).
pubmed: 30595436 pmcid: 6375733 doi: 10.1016/j.molcel.2018.11.024
Duxin, J. P., Dewar, J. M., Yardimci, H. & Walter, J. C. Repair of a DNA–protein crosslink by replication-coupled proteolysis. Cell 159, 346–357 (2014).
pubmed: 25303529 pmcid: 4229047 doi: 10.1016/j.cell.2014.09.024
Mohni, K. N. et al. HMCES maintains genome integrity by shielding abasic sites in single-strand DNA. Cell 176, 144–153 (2019).
pubmed: 30554877 doi: 10.1016/j.cell.2018.10.055
Semlow, D. R., MacKrell, V. A. & Walter, J. C. The HMCES DNA–protein cross-link functions as an intermediate in DNA interstrand cross-link repair. Nat. Struct. Mol. Biol. 29, 451–462 (2022).
pubmed: 35534579 pmcid: 9949344 doi: 10.1038/s41594-022-00764-0
Semlow, D. R., Zhang, J., Budzowska, M., Drohat, A. C. & Walter, J. C. Replication-dependent unhooking of DNA interstrand cross-links by the NEIL3 glycosylase. Cell 167, 498–511 (2016).
pubmed: 27693351 pmcid: 5237264 doi: 10.1016/j.cell.2016.09.008
Haracska, L. et al. Roles of yeast DNA polymerases δ and ζ and of Rev1 in the bypass of abasic sites. Genes Dev. 15, 945–954 (2001).
pubmed: 11316789 pmcid: 312678 doi: 10.1101/gad.882301
Räschle, M. et al. Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links. Science 348, 1253671 (2015).
pubmed: 25931565 pmcid: 5331883 doi: 10.1126/science.1253671
Sertic, S. et al. Coordinated activity of Y family TLS polymerases and EXO1 protects non-S phase cells from UV-induced cytotoxic lesions. Mol. Cell 70, 34–47 (2018).
pubmed: 29551515 doi: 10.1016/j.molcel.2018.02.017
Schubert, L. et al. SCAI promotes error-free repair of DNA interstrand crosslinks via the Fanconi anemia pathway. EMBO Rep. 23, e53639 (2022).
pubmed: 35156773 pmcid: 8982572 doi: 10.15252/embr.202153639
Yoshimura, A., Kobayashi, Y., Tada, S., Seki, M. & Enomoto, T. WRNIP1 functions upstream of DNA polymerase η in the UV-induced DNA damage response. Biochem. Biophys. Res. Commun. 452, 48–52 (2014).
pubmed: 25139235 doi: 10.1016/j.bbrc.2014.08.043
Yuasa, M. S. et al. A human DNA polymerase η complex containing Rad18, Rad6 and Rev1; proteomic analysis and targeting of the complex to the chromatin-bound fraction of cells undergoing replication fork arrest. Genes Cells 11, 731–744 (2006).
pubmed: 16824193 doi: 10.1111/j.1365-2443.2006.00974.x
Chen, D. et al. BRCA1 deficiency specific base substitution mutagenesis is dependent on translesion synthesis and regulated by 53BP1. Nat. Commun. 13, 226 (2022).
pubmed: 35017534 pmcid: 8752635 doi: 10.1038/s41467-021-27872-7
Guo, C. et al. Mouse Rev1 protein interacts with multiple DNA polymerases involved in translesion DNA synthesis. EMBO J. 22, 6621–6630 (2003).
pubmed: 14657033 pmcid: 291821 doi: 10.1093/emboj/cdg626
Malik, R. et al. Structure and mechanism of B-family DNA polymerase ζ specialized for translesion DNA synthesis. Nat. Struct. Mol. Biol. 27, 913–924 (2020).
pubmed: 32807989 pmcid: 7554088 doi: 10.1038/s41594-020-0476-7
Haracska, L., Prakash, L. & Prakash, S. Role of human DNA polymerase κ as an extender in translesion synthesis. Proc. Natl Acad. Sci. USA 99, 16000–16005 (2002).
pubmed: 12444249 pmcid: 138554 doi: 10.1073/pnas.252524999
Washington, M. T., Johnson, R. E., Prakash, L. & Prakash, S. Human DINB1-encoded DNA polymerase κ is a promiscuous extender of mispaired primer termini. Proc. Natl Acad. Sci. USA 99, 1910–1914 (2002).
pubmed: 11842189 pmcid: 122293 doi: 10.1073/pnas.032594399
Kannouche, P. L., Wing, J. & Lehmann, A. R. Interaction of human DNA polymerase η with monoubiquitinated PCNA: a possible mechanism for the polymerase switch in response to DNA damage. Mol. Cell 14, 491–500 (2004).
pubmed: 15149598 doi: 10.1016/S1097-2765(04)00259-X
Lange, S. S., Wittschieben, J. P. & Wood, R. D. DNA polymerase ζ is required for proliferation of normal mammalian cells. Nucleic Acids Res. 40, 4473–4482 (2012).
pubmed: 22319213 pmcid: 3378892 doi: 10.1093/nar/gks054
Ben Yamin, B. et al. DNA polymerase ζ contributes to heterochromatin replication to prevent genome instability. EMBO J. 40, e104543 (2021).
pubmed: 34533226 pmcid: 8561639 doi: 10.15252/embj.2020104543
Wojtaszek, J. L. et al. A small molecule targeting mutagenic translesion synthesis improves chemotherapy. Cell 178, 152–159 (2019).
pubmed: 31178121 pmcid: 6644000 doi: 10.1016/j.cell.2019.05.028
Yamanaka, K., Chatterjee, N., Hemann, M. T. & Walker, G. C. Inhibition of mutagenic translesion synthesis: a possible strategy for improving chemotherapy? PLoS Genet. 13, e1006842 (2017).
pubmed: 28817566 pmcid: 5560539 doi: 10.1371/journal.pgen.1006842
Yu, D., Chojnowski, G., Rosenthal, M. & Kosinski, J. AlphaPulldown—a python package for protein–protein interaction screens using AlphaFold-Multimer. Bioinformatics 39, btac749 (2023).
pubmed: 36413069 doi: 10.1093/bioinformatics/btac749
Jumper, J. et al. Highly accurate protein structure prediction with AlphaFold. Nature 596, 583–589 (2021).
pubmed: 34265844 pmcid: 8371605 doi: 10.1038/s41586-021-03819-2
Yockey, O. P. et al. Mechanism of error-free DNA replication past lucidin-derived DNA damage by human DNA polymerase κ. Chem. Res. Toxicol. 30, 2023–2032 (2017).
pubmed: 28972744 pmcid: 5696069 doi: 10.1021/acs.chemrestox.7b00227
Zhang, Z. et al. Structure of monoubiquitinated PCNA: implications for DNA polymerase switching and Okazaki fragment maturation. Cell Cycle 11, 2128–2136 (2012).
pubmed: 22592530 pmcid: 3368864 doi: 10.4161/cc.20595
Lebofsky, R., Takahashi, T. & Walter, J. C. DNA replication in nucleus-free Xenopus egg extracts. Methods Mol. Biol. 521, 229–252 (2009).
pubmed: 19563110 doi: 10.1007/978-1-60327-815-7_13
Sparks, J. L. et al. The CMG helicase bypasses DNA–protein cross-links to facilitate their repair. Cell 176, 167–181 (2019).
pubmed: 30595447 doi: 10.1016/j.cell.2018.10.053
Sobeck, A. et al. Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks. Mol. Cell. Biol. 26, 425–437 (2006).
pubmed: 16382135 pmcid: 1346898 doi: 10.1128/MCB.26.2.425-437.2006
Knipscheer, P. et al. The Fanconi anemia pathway promotes replication-dependent DNA interstrand cross-link repair. Science 326, 1698–1701 (2009).
pubmed: 19965384 pmcid: 2909596 doi: 10.1126/science.1182372
Räschle, M. et al. Mechanism of replication-coupled DNA interstrand crosslink repair. Cell 134, 969–980 (2008).
pubmed: 18805090 pmcid: 2748255 doi: 10.1016/j.cell.2008.08.030
Sparks, J. & Walter, J. C. Extracts for analysis of DNA replication in a nucleus-free system. Cold Spring Harb. Protoc. 2019, prot097154 (2019).
doi: 10.1101/pdb.prot097154
Evans, R. et al. Protein complex prediction with AlphaFold-Multimer. Preprint at bioRxiv https://doi.org/10.1101/2021.10.04.463034 (2022).
Varadi, M. et al. AlphaFold Protein Structure Database: massively expanding the structural coverage of protein-sequence space with high-accuracy models. Nucleic Acids Res. 50, D439–D444 (2022).
pubmed: 34791371 doi: 10.1093/nar/gkab1061
Goddard, T. D. et al. UCSF ChimeraX: meeting modern challenges in visualization and analysis. Protein Sci. 27, 14–25 (2018).
pubmed: 28710774 doi: 10.1002/pro.3235
Pettersen, E. F. et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021).
pubmed: 32881101 doi: 10.1002/pro.3943
Lancey, C. et al. Structure of the processive human Pol δ holoenzyme. Nat. Commun. 11, 1109 (2020).
pubmed: 32111820 pmcid: 7048817 doi: 10.1038/s41467-020-14898-6
Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367–1372 (2008).
pubmed: 19029910 doi: 10.1038/nbt.1511
Cox, J. et al. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol. Cell. Proteomics 13, 2513–2526 (2014).
pubmed: 24942700 pmcid: 4159666 doi: 10.1074/mcp.M113.031591
Tyanova, S. et al. The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat. Methods 13, 731–740 (2016).
pubmed: 27348712 doi: 10.1038/nmeth.3901
Li, W. et al. MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens. Genome Biol. 15, 554 (2014).
pubmed: 25476604 pmcid: 4290824 doi: 10.1186/s13059-014-0554-4
Ritchie, M. E. et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 43, e47 (2015).
pubmed: 25605792 pmcid: 4402510 doi: 10.1093/nar/gkv007
Richter, M. F. et al. Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity. Nat. Biotechnol. 38, 883–891 (2020).
pubmed: 32433547 pmcid: 7357821 doi: 10.1038/s41587-020-0453-z
Perez-Riverol, Y. et al. The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences. Nucleic Acids Res. 50, D543–D552 (2022).
pubmed: 34723319 doi: 10.1093/nar/gkab1038

Auteurs

Selene Sellés-Baiget (S)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Sara M Ambjørn (SM)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Alberto Carli (A)

Center for Chromosome Stability, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Ivo A Hendriks (IA)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Irene Gallina (I)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
Department of Molecular Medicine, University of Padua, Padua, Italy.

Norman E Davey (NE)

Division of Cancer Biology, The Institute of Cancer Research, London, UK.

Bente Benedict (B)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Alessandra Zarantonello (A)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Sampath A Gadi (SA)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Bob Meeusen (B)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Emil P T Hertz (EPT)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Laura Slappendel (L)

Department of Health Sciences and Technology, ETH Zurich, Zurich, Switzerland.

Daniel Semlow (D)

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, USA.

Shana Sturla (S)

Department of Health Sciences and Technology, ETH Zurich, Zurich, Switzerland.

Michael L Nielsen (ML)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Jakob Nilsson (J)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.

Thomas C R Miller (TCR)

Center for Chromosome Stability, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Julien P Duxin (JP)

Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark. Julien.duxin@bric.ku.dk.
Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. Julien.duxin@bric.ku.dk.

Classifications MeSH