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.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Journal of Experimental & Clinical Cancer Research Open Access 14 March 2022
Experimental & Molecular Medicine Open Access 18 December 2020
Nature Communications Open Access 14 July 2020
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Lengronne, A. et al. Establishment of sister chromatid cohesion at the S.cerevisiae replication fork. Mol. Cell 23, 787–799 (2006)
Gullerova, M. & Proudfoot, N. J. Cohesin complex promotes transcriptional termination between convergent genes in S.pombe. Cell 132, 983–995 (2008)
Rubio, E. D. et al. CTCF physically links cohesin to chromatin. Proc. Natl Acad. Sci. USA 105, 8309–8314 (2008)
Parelho, V. et al. Cohesins functionally associate with CTCF on mammalian chromosome arms. Cell 132, 422–433 (2008)
Wendt, K. S. et al. Cohesin mediates transcriptional insulation by CCCTC-binding factor. Nature 451, 796–801 (2008)
Stedman, W. et al. Cohesins localize with CTCF at the KSHV latency control region and at cellular c-myc and H19/Igf2 insulators. EMBO J. 27, 654–666 (2008)
Bermudez, V. P. et al. The alternative Ctf18-Dcc1-Ctf8-replication factor C complex required for sister chromatid cohesion loads proliferating cell nuclear antigen onto DNA. Proc. Natl Acad. Sci. USA 100, 10237–10242 (2003)
Zhang, J. et al. Acetylation of Smc3 by Eco1 is required for S phase sister chromatid cohesion in both human and yeast. Mol. Cell 31, 143–151 (2008)
Unal, E. et al. A molecular determinant for the establishment of sister chromatid cohesion. Science 321, 566–569 (2008)
Ben-Shahar, T. R. et al. Eco1-dependent cohesin acetylation during establishment of sister chromatid cohesion. Science 321, 563–566 (2008)
Vega, H. et al. Roberts syndrome is caused by mutations in ESCO2, a human homolog of yeast ECO1 that is essential for the establishment of sister chromatid cohesion. Nature Genet. 37, 468–470 (2005)
Gandhi, R., Gillespie, P. J. & Hirano, T. Human Wapl is a cohesin-binding protein that promotes sister-chromatid resolution in mitotic prophase. Curr. Biol. 16, 2406–2417 (2006)
Kueng, S. et al. Wapl controls the dynamic association of cohesin with chromatin. Cell 127, 955–967 (2006)
Lengronne, A. et al. Cohesin relocation from sites of chromosomal loading to places of convergent transcription. Nature 430, 573–578 (2004)
Walter, J. & Newport, J. W. Regulation of replicon size in Xenopus egg extracts. Science 275, 993–995 (1997)
Deng, Y., Chan, S. S. & Chang, S. Telomere dysfunction and tumour suppression: the senescence connection. Nature Rev. Cancer 8, 450–458 (2008)
Wang, X. et al. Rad17 phosphorylation is required for claspin recruitment and Chk1 activation in response to replication stress. Mol. Cell 23, 331–341 (2006)
Heller, R. C. & Marians, K. J. Replisome assembly and the direct restart of stalled replication forks. Nature Rev. Mol. Cell Biol. 7, 932–943 (2006)
Courbet, S. et al. Replication fork movement sets chromatin loop size and origin choice in mammalian cells. Nature 455, 557–560 (2008)
Shay, J. W., Pereira-Smith, O. M. & Wright, W. E. A role for both RB and p53 in the regulation of human cellular senescence. Exp. Cell Res. 196, 33–39 (1991)
Takahashi, T. S., Basu, A., Bermudez, V., Hurwitz, J. & Walter, J. C. Cdc7-Drf1 kinase links chromosome cohesion to the initiation of DNA replication in Xenopus egg extracts. Genes Dev. 22, 1894–1905 (2008)
Moldovan, G. L., Pfander, B. & Jentsch, S. PCNA controls establishment of sister chromatid cohesion during S phase. Mol. Cell 23, 723–732 (2006)
Tanaka, K., Hao, Z., Kai, M. & Okayama, H. Establishment and maintenance of sister chromatid cohesion in fission yeast by a unique mechanism. EMBO J. 20, 5779–5790 (2001)
Losada, A., Yokochi, T. & Hirano, T. Functional contribution of Pds5 to cohesin-mediated cohesion in human cells and Xenopus egg extracts. J. Cell Sci. 118, 2133–2141 (2005)
Rowland, B. D. et al. Building sister chromatid cohesion: Smc3 acetylation counteracts an antiestablishment activity. Mol. Cell 33, 763–774 (2009)
Sutani, T., Kawaguchi, T., Kanno, R., Itoh, T. & Shirahige, K. Budding yeast Wpl1(Rad61)-Pds5 complex counteracts sister chromatid cohesion-establishing reaction. Curr. Biol. 19, 492–497 (2009)
Mc Intyre, J. et al. In vivo analysis of cohesin architecture using FRET in the budding yeast Saccharomyces cerevisiae. EMBO J. 26, 3783–3793 (2007)
Sakai, A., Hizume, K., Sutani, T., Takeyasu, K. & Yanagida, M. Condensin but not cohesin SMC heterodimer induces DNA reannealing through protein–protein assembly. EMBO J. 22, 2764–2775 (2003)
Branzei, D. & Foiani, M. Regulation of DNA repair throughout the cell cycle. Nature Rev. Mol. Cell Biol. 9, 297–308 (2008)
Jackson, D. A. & Pombo, A. Replicon clusters are stable units of chromosome structure: evidence that nuclear organization contributes to the efficient activation and propagation of S phase in human cells. J. Cell Biol. 140, 1285–1295 (1998)
Papi, M., Berdougo, E., Randall, C. L., Ganguly, S. & Jallepalli, P. V. Multiple roles for separase auto-cleavage during the G2/M transition. Nature Cell Biol. 7, 1029–1035 (2005)
Burkard, M. E. et al. Chemical genetics reveals the requirement for Polo-like kinase 1 activity in positioning RhoA and triggering cytokinesis in human cells. Proc. Natl Acad. Sci. USA 104, 4383–4388 (2007)
Serrano, M., Lin, A. W., McCurrach, M. E., Beach, D. & Lowe, S. W. Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88, 593–602 (1997)
Méndez, J. & Stillman, B. Chromatin association of human origin recognition complex, Cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis. Mol. Cell. Biol. 20, 8602–8612 (2000)
Ishihara, K., Oshimura, M. & Nakao, M. CTCF-dependent chromatin insulator is linked to epigenetic remodeling. Mol. Cell 23, 733–742 (2006)
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.
About this article
Cite this article
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
This article is cited by
Journal of Experimental & Clinical Cancer Research (2022)
Nature Communications (2020)
Experimental & Molecular Medicine (2020)
Nature Reviews Cancer (2020)