The Prereplication Complex Recruits XEco2 to Chromatin to Promote Cohesin Acetylation in Xenopus Egg Extracts

Higashi, Torahiko L.; Ikeda, Megumi; Tanaka, Hiroshi; Nakagawa, Takuro; Bando, Masashige; Shirahige, Katsuhiko; Kubota, Yumiko; Takisawa, Haruhiko; Masukata, Hisao; Takahashi, Tatsuro

doi:10.1016/j.cub.2012.04.013

Cited by 55 publications

(69 citation statements)

References 44 publications

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“…Although no structure has been assigned to the N-terminal region of XEco2/Esco2, it is possible that the loss of chromatin binding in the double deletion is due to improper folding of the protein. However, our results are consistent with findings by others attributing chromatin binding to the poorly conserved N-terminal region of the protein (8,31). Again, we noted that Sororin recruitment to chromatin was dependent on Smc3 acetylation; mutant versions of XEco2 that were compromised for Smc3 acetylation did not promote Sororin recruitment (Fig.…”

Section: Xeco2supporting

confidence: 93%

“…Although actinomycin D, an intercalating agent known to inhibit primase (30), did reduce Smc3 acetylation as reported previously (14), this effect may be indirect because we noted that recruitment of both Smc3 and Pds5 was reduced with this treatment. Consistent with previous studies (10,12,31), addition of the replication-licensing factor geminin blocked recruitment of both cohesin and XEco2. This resulted in a decrease in detectable acetylated cohesin, in keeping with the model that cohesin must be loaded onto chromatin to be acetylated, as seen in other systems (27,32).…”

Section: Xeco2supporting

confidence: 91%

“…Our data suggest that XEco2 is likely recruited through interaction of conserved motifs in its N terminus with a chromatin-associated partner. The identity of the interacting partner is of great interest: it is unlikely to be cohesin itself, because XEco2 is recruited in a cohesin-independent manner and does not interact with cohesin in vitro (9,31). Interestingly, it was shown recently that the recruitment of XEco2 to chromatin, like cohesin, is dependent on the early steps in assembly of the replication machinery (31).…”

Section: Discussionmentioning

confidence: 99%

“…The identity of the interacting partner is of great interest: it is unlikely to be cohesin itself, because XEco2 is recruited in a cohesin-independent manner and does not interact with cohesin in vitro (9,31). Interestingly, it was shown recently that the recruitment of XEco2 to chromatin, like cohesin, is dependent on the early steps in assembly of the replication machinery (31). Thus the replication machinery controls XEco2 function at multiple levels: first through recruitment and subsequently through interaction with PCNA.…”

Section: Discussionmentioning

confidence: 99%

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Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

Song

Lafont

Chen

et al. 2012

Journal of Biological Chemistry

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Background:The conversion of cohesin to an active form requires DNA replication and acetylation of the Smc3 subunit of the complex by Eco acetyltransferases. Results: Cohesin acetylation occurs both when replication is blocked and after replication is complete but only ensures cohesion in association with the replication machinery. Conclusion: The context of cohesin acetylation is critical to cohesion establishment. Significance: Acetylation and cohesion establishment are regulated differently in yeast and vertebrates.

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Section: Xeco2supporting

confidence: 93%

Section: Xeco2supporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

Song

Lafont

Chen

et al. 2012

Journal of Biological Chemistry

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“…We examined the salt extraction properties of Xcohesin following DNA replication. It was recently reported that X-Smc3 subunit of cohesin becomes acetylated after being loaded onto pre-RCs, and this modification did not require replication (24). The acetylated cohesin formed under these conditions was salt labile, but following replication, it was rendered salt-stable.…”

Section: Resultsmentioning

confidence: 96%

In vitro loading of human cohesin on DNA by the human Scc2-Scc4 loader complex

Bermudez

Farina

Higashi

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The loading of cohesin onto chromatin requires the heterodimeric complex sister chromatid cohesion (Scc)2 and Scc4 (Scc2/4), which is highly conserved in all species. Here, we describe the purification of the human (h)-Scc2/4 and show that it interacts with hcohesin and the heterodimeric Smc1-Smc3 complex but not with the Smc1 or Smc3 subunit alone. We demonstrate that both h-Scc2/4 and h-cohesin are loaded onto dsDNA containing the prereplication complex (pre-RC) generated in vitro by Xenopus high-speed soluble extracts. The addition of geminin, which blocks pre-RC formation, prevents the loading of Scc2/4 and cohesin. Xenopus extracts depleted of endogenous Scc2/4 with specific antibodies, although able to form pre-RCs, did not support cohesin loading unless supplemented with purified h-Scc2/4. The results presented here indicate that the Xenopus or h-Scc2/4 complex supports the loading of Xenopus and/or h-cohesin onto pre-RCs formed by Xenopus high-speed extracts. We show that cohesin loaded onto pre-RCs either by h-Scc2/4 and/or the Xenopus complex was dissociated from chromatin by low salt extraction, similar to cohesin loaded onto chromatin in G 1 by HeLa cells in vivo. Replication of cohesin-loaded DNA, both in vitro and in vivo, markedly increased the stability of cohesin associated with DNA. Collectively, these in vitro findings partly recapitulate the in vivo pathway by which sister chromatids are linked together, leading to cohesion.cohesin stability | cell-cycle | cohesion establishment | sister chromatid cohesion N ewly replicated chromosomes are held together until their separation and equal distribution to daughter cells during cell division. Their association is mediated by cohesin, a foursubunit complex comprising (i) Rad21/sister chromatid cohesion (Scc)1/multiple chloroplast division (Mcd)1, (ii) structural maintenance of chromosomes (Smc)1, (iii) Smc3, and (iv) Scc3/ (stromal antigen) STAG/SA that stably links the sister chromatids together in S and G 2 (1). Smc1 and Smc3 are elongated coiled-coil proteins, each of which folds onto itself to form a globular ATPase head domain through the juxtaposition of the N and C terminus with a hinge domain at the other end. Smc1 and Smc3 interact through their hinge domains to form a heterodimer. The kleisin subunit, Scc1, associates with the head domains of Smc1 and Smc3 to stabilize their interaction and recruits the Scc3/SA subunit. The Smc1-Smc3-Scc1 complex forms a ring structure with an internal diameter of 40 nm, large enough to tether two chromatids (embrace model) (2). Although other models have been proposed to explain how the cohesin ring leads to the association of sister chromatids, substantial evidence supporting the "embrace model" has accumulated (3).Sister chromatid cohesion is a multistep process that includes cohesin loading onto chromatin and the establishment of cohesion during replication (3). Although cohesin loading required for cohesion occurs before the S phase, the timing of its chromosome association differs among species. In ...

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CyclinD1 Down‐Regulation and Increased Apoptosis Are Common Features of Cohesinopathies

Fazio

Gaston‐Massuet

Bettini

et al. 2015

Journal Cellular Physiology

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Genetic variants within components of the cohesin complex (NIPBL, SMC1A, SMC3, RAD21, PDS5, ESCO2, HDAC8) are believed to be responsible for a spectrum of human syndromes known as "cohesinopathies" that includes Cornelia de Lange Syndrome (CdLS). CdLS is a multiple malformation syndrome affecting almost any organ and causing severe developmental delay. Cohesinopathies seem to be caused by dysregulation of specific developmental pathways downstream of mutations in cohesin components. However, it is still unclear how mutations in different components of the cohesin complex affect the output of gene regulation. In this study, zebrafish embryos and SMC1A-mutated patient-derived fibroblasts were used to analyze abnormalities induced by SMC1A loss of function. We show that the knockdown of smc1a in zebrafish impairs neural development, increases apoptosis, and specifically down-regulates Ccnd1 levels. The same down-regulation of cohesin targets is observed in SMC1A-mutated patient fibroblasts. Previously, we have demonstrated that haploinsufficiency of NIPBL produces similar effects in zebrafish and in patients fibroblasts indicating a possible common feature for neurological defects and mental retardation in cohesinopathies. Interestingly, expression analysis of Smc1a and Nipbl in developing mouse embryos reveals a specific pattern in the hindbrain, suggesting a role for cohesins in neural development in vertebrates.

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The Prereplication Complex Recruits XEco2 to Chromatin to Promote Cohesin Acetylation in Xenopus Egg Extracts

Abstract: Our results demonstrate that pre-RC formation regulates chromatin association of XEco2 in Xenopus egg extracts. We propose that this reaction is critical to acetylate cohesin, whose DNA binding is subsequently stabilized by DNA replication.

Cited by 55 publications

References 44 publications

Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

Cohesin Acetylation Promotes Sister Chromatid Cohesion Only in Association with the Replication Machinery

In vitro loading of human cohesin on DNA by the human Scc2-Scc4 loader complex

CyclinD1 Down‐Regulation and Increased Apoptosis Are Common Features of Cohesinopathies

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