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Published online 7 August 2000. doi:10.1083/jcb.150.3.405
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© The Rockefeller University Press, 0021-9525/2000/8/405/ $5.00
The Journal of Cell Biology, Volume 150, Number 3, August 7, 2000 405-416


Original Article

Identification and Characterization of SA/Scc3p Subunits in the Xenopus and Human Cohesin Complexes

Ana Losadaa, Tomoki Yokochia, Ryuji Kobayashia, and Tatsuya Hiranoa
a Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724

Correspondence to: Tatsuya Hirano, Cold Spring Harbor Laboratory, 1 Bungtown Road, P.O. Box 100, Cold Spring Harbor, NY 11724. Tel:(516) 367-8370 Fax:(516) 367-8815 E-mail:hirano{at}cshl.org.

A multisubunit protein complex, termed cohesin, plays an essential role in sister chromatid cohesion in yeast and in Xenopus laevis cell-free extracts. We report here that two distinct cohesin complexes exist in Xenopus egg extracts. A 14S complex (x-cohesinSA1) contains XSMC1, XSMC3, XRAD21, and a newly identified subunit, XSA1. In a second 12.5S complex (x-cohesinSA2), XSMC1, XSMC3, and XRAD21 associate with a different subunit, XSA2. Both XSA1 and XSA2 belong to the SA family of mammalian proteins and exhibit similarity to Scc3p, a recently identified component of yeast cohesin. In Xenopus egg extracts, x-cohesinSA1 is predominant, whereas x-cohesinSA2 constitutes only a very minor population. Human cells have a similar pair of cohesin complexes, but the SA2-type is the dominant form in somatic tissue culture cells. Immunolocalization experiments suggest that chromatin association of cohesinSA1 and cohesinSA2 may be differentially regulated. Dissociation of x-cohesinSA1 from chromatin correlates with phosphorylation of XSA1 in the cell-free extracts. Purified cdc2-cyclin B can phosphorylate XSA1 in vitro and reduce the ability of x-cohesinSA1 to bind to DNA or chromatin. These results shed light on the mechanism by which sister chromatid cohesion is partially dissolved in early mitosis, far before the onset of anaphase, in vertebrate cells.

Key Words: Xenopus egg extract, sister chromatid cohesion, SMC proteins, chromosome structure, mitosis


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