Scc2 couples replication licensing to sister chromatid cohesion in Xenopus egg extracts

Gillespie, P. J., Hirano, T. (2004) Scc2 couples replication licensing to sister chromatid cohesion in Xenopus egg extracts. Curr Biol, 14 (17). pp. 1598-603. ISSN 0960-9822 (Print)

URL: http://www.sciencedirect.com/science/journal/09609...
DOI: 10.1016/j.cub.2004.07.053

Abstract

The cohesin complex is a central player in sister chromatid cohesion, a process that ensures the faithful segregation of chromosomes in mitosis and meiosis. Previous genetic studies in yeast show that Scc2/Mis4, a HEAT-repeat-containing protein, is required for the loading of cohesin onto chromatin. In this study, we have identified two isoforms of Scc2 in humans and Xenopus (termed Scc2A and Scc2B), which are encoded by a single gene but have different carboxyl termini created by alternative splicing. Both Scc2A and Scc2B bind to chromatin concomitant with cohesin during DNA replication in Xenopus egg extracts. Simultaneous immunodepletion of Scc2A and Scc2B from the extracts impairs the association of cohesin with chromatin, leading to severe defects in sister chromatid pairing in the subsequent mitosis. The loading of Scc2 onto chromatin is inhibited in extracts treated with geminin but not with p21(CIP1), suggesting that this step depends on replication licensing but not on the initiation of DNA replication. Upon mitotic entry, Scc2 is removed from chromatin through a mechanism that requires cdc2 but not aurora B or polo-like kinase. Our results suggest that vertebrate Scc2 couples replication licensing to sister chromatid cohesion by facilitating the loading of cohesin onto chromatin.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Animals Cell Cycle Proteins Cell Extracts Chromatids metabolism physiology Chromatography Gel Chromosomal Proteins Non-Histone genetics metabolism DNA Replication physiology DNA, Complementary genetics Fungal Proteins Immunoblotting Immunoprecipitation Mitosis physiology Molecular Sequence Data Nuclear Proteins metabolism Ovum cytology metabolism Protein Isoforms Sequence Alignment Sequence Analysis DNA Xenopus genetics metabolism Xenopus Proteins genetics metabolism
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > chromatid
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > cohesin complex
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Depositing User: CSHL Librarian
Date Deposited: 03 Feb 2012 16:51
Last Modified: 03 Feb 2012 16:51
URI: http://repository.cshl.edu/id/eprint/22373

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