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Replication forks proteomics reveals ATM-dependent rewiring of broken forks to promote restart

ABSTRACT: The fidelity of DNA replication is governed by a network of DNA repair mechanisms. Defects in replication fork protection can fuel genome instability in cancer, while also creating cancer-specific vulnerabilities. Here, we provide a comprehensive proteomic characterization of the replication fork response to topoisomerase 1 inhibition, a widely used mainstay chemotherapy. We reveal profound changes in the fork proteome and chromatin environment in response to seDSBs and topological stress, and classify fork repair factors according to their specificity for broken and stalled forks. These distinct fork responses also oppositely affect nucleosome occupancy and nuclear membrane interactions. ATM inhibition dramatically rewired the fork breakage response, revealing that ATM promotes HR-dependent fork restart by concomitantly promoting DNA-end resection and suppressing DSB associated protein ubiquitination and NHEJ. Together, this collection of damaged fork proteomes provides an ample resource to understand fork repair mechanisms and identify druggable targets and novel cancer vulnerabilities.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Diploid Cell

SUBMITTER: Georg Kustatscher

LAB HEAD: Juri Rappsilber

PROVIDER: PXD018092 | Pride | 2021-01-15

REPOSITORIES: Pride

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			Action	DRS
	110428_04_Orbi2_FLA_CO_185_AG_Constance_SampleB_F5.raw	Raw
	110428_05_Orbi2_FLA_CO_185_AG_Constance_SampleB_F6.raw	Raw
	110428_06_Orbi2_FLA_CO_185_AG_Constance_SampleB_F10.raw	Raw
	110428_07_Orbi2_FLA_CO_185_AG_Constance_SampleB_F11.raw	Raw
	110428_08_Orbi2_FLA_CO_185_AG_Constance_SampleB_F21.raw	Raw

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