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

Markus Räschle; Godelieve Smeenk; Rebecca K Hansen; Tikira Temu; Yasuyoshi Oka; Marco Y Hein; Nagarjuna Nagaraj; David T Long; Johannes C Walter; Kay Hofmann; Zuzana Storchova; Jürgen Cox; Simon Bekker-Jensen; Niels Mailand; Matthias Mann

doi:10.1126/science.1253671

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

Markus Räschle, Godelieve Smeenk, Rebecca K Hansen, Tikira Temu, Yasuyoshi Oka, Marco Y Hein, Nagarjuna Nagaraj, David T Long, Johannes C Walter, Kay Hofmann, Zuzana Storchova, Jürgen Cox, Simon Bekker-Jensen, Niels Mailand, Matthias Mann

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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.

Original language	English
Article number	1253671
Journal	Science (New York, N.Y.)
Volume	348
Issue number	6234
Pages (from-to)	539-
Number of pages	10
ISSN	0036-8075
DOIs	https://doi.org/10.1126/science.1253671
Publication status	Published - 1 May 2015

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Räschle, M., Smeenk, G., Hansen, R. K., Temu, T., Oka, Y., Hein, M. Y., Nagaraj, N., Long, D. T., Walter, J. C., Hofmann, K., Storchova, Z., Cox, J., Bekker-Jensen, S., Mailand, N., & Mann, M. (2015). Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links. Science (New York, N.Y.), 348(6234), 539-. Article 1253671. https://doi.org/10.1126/science.1253671

Räschle, M, Smeenk, G, Hansen, RK, Temu, T, Oka, Y, Hein, MY, Nagaraj, N, Long, DT, Walter, JC, Hofmann, K, Storchova, Z, Cox, J, Bekker-Jensen, S , Mailand, N & Mann, M 2015, 'Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links', Science (New York, N.Y.), vol. 348, no. 6234, 1253671, pp. 539-. https://doi.org/10.1126/science.1253671

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title = "Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links",

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.",

keywords = "Animals, Chromatin, DNA Damage, DNA Repair, DNA Repair Enzymes, DNA Replication, DNA-Binding Proteins, Mass Spectrometry, Proteomics, RNA-Binding Proteins, Xenopus",

author = "Markus R{\"a}schle and Godelieve Smeenk and Hansen, {Rebecca K} and Tikira Temu and Yasuyoshi Oka and Hein, {Marco Y} and Nagarjuna Nagaraj and Long, {David T} and Walter, {Johannes C} and Kay Hofmann and Zuzana Storchova and J{\"u}rgen Cox and Simon Bekker-Jensen and Niels Mailand and Matthias Mann",

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T1 - Proteomics reveals dynamic assembly of repair complexes during bypass of DNA cross-links

AU - Räschle, Markus

AU - Smeenk, Godelieve

AU - Hansen, Rebecca K

AU - Temu, Tikira

AU - Oka, Yasuyoshi

AU - Hein, Marco Y

AU - Nagaraj, Nagarjuna

AU - Long, David T

AU - Walter, Johannes C

AU - Hofmann, Kay

AU - Storchova, Zuzana

AU - Cox, Jürgen

AU - Bekker-Jensen, Simon

AU - Mailand, Niels

AU - Mann, Matthias

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N2 - 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.

AB - 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.

KW - Animals

KW - Chromatin

KW - DNA Damage

KW - DNA Repair

KW - DNA Repair Enzymes

KW - DNA Replication

KW - DNA-Binding Proteins

KW - Mass Spectrometry

KW - Proteomics

KW - RNA-Binding Proteins

KW - Xenopus

U2 - 10.1126/science.1253671

DO - 10.1126/science.1253671

M3 - Journal article

C2 - 25931565

SN - 0036-8075

VL - 348

SP - 539-

JO - Science (New York, N.Y.)

JF - Science (New York, N.Y.)

IS - 6234

M1 - 1253671

ER -

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

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