Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links

Markus Räschle(Max Planck Institute of Biochemistry), Godelieve Smeenk(Novo Nordisk Foundation), Rebecca K. Hansen(Novo Nordisk Foundation), Tikira Temu(Max Planck Institute of Biochemistry), Yasuyoshi Oka(Novo Nordisk Foundation), Marco Y. Hein(Max Planck Institute of Biochemistry), Nagarjuna Nagaraj(Max Planck Institute of Biochemistry), David T. Long(Howard Hughes Medical Institute), Johannes C. Walter(Howard Hughes Medical Institute), Kay Hofmann(University of Cologne), Zuzana Štorchová(Max Planck Institute of Biochemistry), Jürgen Cox(Max Planck Institute of Biochemistry), Simon Bekker‐Jensen(Novo Nordisk Foundation), Niels Mailand(Novo Nordisk Foundation), Matthias Mann(Novo Nordisk Foundation)
Science
April 30, 2015
10.1126/science.1253671
Cited by 240Open Access
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Abstract

DNA interstrand cross-links (ICLs) block replication fork progression by inhibiting DNA strand separation. Repair of ICLs requires sequential incisions, translesion DNA synthesis, and homologous recombination, but the full set of factors involved in these transactions remains unknown. We devised a technique called chromatin mass spectrometry (CHROMASS) to study protein recruitment dynamics during perturbed DNA replication in Xenopus egg extracts. Using CHROMASS, we systematically monitored protein assembly and disassembly on ICL-containing chromatin. Among numerous prospective DNA repair factors, we identified SLF1 and SLF2, which form a complex with RAD18 and together define a pathway that suppresses genome instability by recruiting the SMC5/6 cohesion complex to DNA lesions. Our study provides a global analysis of an entire DNA repair pathway and reveals the mechanism of SMC5/6 relocalization to damaged DNA in vertebrate cells.

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