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Cohesin acetylation speeds the replication fork

Abstract

Cohesin not only links sister chromatids but also inhibits the transcriptional machinery’s interaction with and movement along chromatin1,2,3,4,5,6. In contrast, replication forks must traverse such cohesin-associated obstructions to duplicate the entire genome in S phase. How this occurs is unknown. Through single-molecule analysis, we demonstrate that the replication factor C (RFC)–CTF18 clamp loader (RFCCTF18)1,7 controls the velocity, spacing and restart activity of replication forks in human cells and is required for robust acetylation of cohesin’s SMC3 subunit and sister chromatid cohesion. Unexpectedly, we discovered that cohesin acetylation itself is a central determinant of fork processivity, as slow-moving replication forks were found in cells lacking the Eco1-related acetyltransferases ESCO1 or ESCO2 (refs 8–10) (including those derived from Roberts’ syndrome patients, in whom ESCO2 is biallelically mutated11) and in cells expressing a form of SMC3 that cannot be acetylated. This defect was a consequence of cohesin’s hyperstable interaction with two regulatory cofactors, WAPL and PDS5A (refs 12, 13); removal of either cofactor allowed forks to progress rapidly without ESCO1, ESCO2, or RFCCTF18. Our results show a novel mechanism for clamp-loader-dependent fork progression, mediated by the post-translational modification and structural remodelling of the cohesin ring. Loss of this regulatory mechanism leads to the spontaneous accrual of DNA damage and may contribute to the abnormalities of the Roberts’ syndrome cohesinopathy.

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Figure 1: The RFC CTF18 complex is required to prevent accumulation of endogenous DNA damage and terminal senescence.
Figure 2: RFC CTF18 controls global replication dynamics and promotes fork reactivation after genotoxic stress.
Figure 3: SMC3 acetylation and replication fork progression are interdependent.
Figure 4: Processive fork movement requires acetylation-mediated dissociation of PDS5A and WAPL.

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Acknowledgements

We thank D. Galloway, J. Hurwitz, J. Petrini, H. Nakao and H. Zou for reagents, and S. Keeney, K. Marians and J. Petrini for discussions and reading of the manuscript. We thank A. Viale, M. Hassimi and the MSKCC Genomics Core Laboratory for assistance with microarray experiments. This work was supported by a grant from the National Institutes of Health and a Pew Scholar in the Biochemical Sciences award to P.V.J.

Author Contributions M.-E.T. and P.V.J. designed experiments, M.-E.T., R.S. and S.R. performed experiments, J.Q. contributed reagents, M.-E.T., R.S., S.R. and P.V.J. analysed the data, and M.-E.T. and P.V.J. wrote the paper.

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Correspondence to Prasad V. Jallepalli.

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Terret, ME., Sherwood, R., Rahman, S. et al. Cohesin acetylation speeds the replication fork. Nature 462, 231–234 (2009). https://doi.org/10.1038/nature08550

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