Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Proc Natl Acad Sci U S A
2012 Jun 12;10924:9366-71. doi: 10.1073/pnas.1206840109.
Show Gene links
Show Anatomy links
In vitro loading of human cohesin on DNA by the human Scc2-Scc4 loader complex.
Bermudez VP, Farina A, Higashi TL, Du F, Tappin I, Takahashi TS, Hurwitz J.
???displayArticle.abstract???
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 h-cohesin 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.
Boos,
Activation of the replicative DNA helicase: breaking up is hard to do.
2012, Pubmed
Boos,
Activation of the replicative DNA helicase: breaking up is hard to do.
2012,
Pubmed Ciosk,
Cohesin's binding to chromosomes depends on a separate complex consisting of Scc2 and Scc4 proteins.
2000,
Pubmed D'Ambrosio,
Identification of cis-acting sites for condensin loading onto budding yeast chromosomes.
2008,
Pubmed Dorsett,
Cohesin: genomic insights into controlling gene transcription and development.
2011,
Pubmed Edwards,
MCM2-7 complexes bind chromatin in a distributed pattern surrounding the origin recognition complex in Xenopus egg extracts.
2002,
Pubmed
,
Xenbase Gerlich,
Live-cell imaging reveals a stable cohesin-chromatin interaction after but not before DNA replication.
2006,
Pubmed Gillespie,
Scc2 couples replication licensing to sister chromatid cohesion in Xenopus egg extracts.
2004,
Pubmed
,
Xenbase Gruber,
Chromosomal cohesin forms a ring.
2003,
Pubmed Higashi,
The prereplication complex recruits XEco2 to chromatin to promote cohesin acetylation in Xenopus egg extracts.
2012,
Pubmed
,
Xenbase Hu,
ATP hydrolysis is required for relocating cohesin from sites occupied by its Scc2/4 loading complex.
2011,
Pubmed Kim,
Genome-wide reinforcement of cohesin binding at pre-existing cohesin sites in response to ionizing radiation in human cells.
2010,
Pubmed Lengronne,
Cohesin relocation from sites of chromosomal loading to places of convergent transcription.
2004,
Pubmed Mannini,
The expanding universe of cohesin functions: a new genome stability caretaker involved in human disease and cancer.
2010,
Pubmed Nasmyth,
Cohesin: a catenase with separate entry and exit gates?
2011,
Pubmed Nasmyth,
Cohesin: its roles and mechanisms.
2009,
Pubmed Nishiyama,
Sororin mediates sister chromatid cohesion by antagonizing Wapl.
2010,
Pubmed
,
Xenbase Nonaka,
Recruitment of cohesin to heterochromatic regions by Swi6/HP1 in fission yeast.
2002,
Pubmed Onn,
In vitro assembly of physiological cohesin/DNA complexes.
2011,
Pubmed Onn,
Sister chromatid cohesion: a simple concept with a complex reality.
2008,
Pubmed Pacek,
A requirement for MCM7 and Cdc45 in chromosome unwinding during eukaryotic DNA replication.
2004,
Pubmed
,
Xenbase Rowles,
Changes in association of the Xenopus origin recognition complex with chromatin on licensing of replication origins.
1999,
Pubmed
,
Xenbase Siegel,
Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases.
1966,
Pubmed Sjögren,
S-phase and DNA damage activated establishment of sister chromatid cohesion--importance for DNA repair.
2010,
Pubmed Strachan,
Cornelia de Lange Syndrome and the link between chromosomal function, DNA repair and developmental gene regulation.
2005,
Pubmed Ström,
Postreplicative recruitment of cohesin to double-strand breaks is required for DNA repair.
2004,
Pubmed Takahashi,
Cdc7-Drf1 kinase links chromosome cohesion to the initiation of DNA replication in Xenopus egg extracts.
2008,
Pubmed
,
Xenbase Takahashi,
Recruitment of Xenopus Scc2 and cohesin to chromatin requires the pre-replication complex.
2004,
Pubmed
,
Xenbase Uhlmann,
A matter of choice: the establishment of sister chromatid cohesion.
2009,
Pubmed Walter,
Regulated chromosomal DNA replication in the absence of a nucleus.
1998,
Pubmed
,
Xenbase Watrin,
Human Scc4 is required for cohesin binding to chromatin, sister-chromatid cohesion, and mitotic progression.
2006,
Pubmed
,
Xenbase Wendt,
How cohesin and CTCF cooperate in regulating gene expression.
2009,
Pubmed
,
Xenbase
