Letter abstract
Nature Cell Biology 6, 991 - 996 (2004)
Published online: 26 September 2004 | doi:10.1038/ncb1177
Recruitment of Xenopus Scc2 and cohesin to chromatin requires the pre-replication complex
Tatsuro S. Takahashi1, Pannyun Yiu1, Michael F. Chou2, Steven Gygi2 & Johannes C. Walter1
Cohesin is a multi-subunit, ring-shaped protein complex that holds sister chromatids together from the time of their synthesis in S phase until they are segregated in anaphase1, 2, 3, 4, 5, 6, 7. In yeast, the loading of cohesin onto chromosomes requires the Scc2 protein8, 9, 10. In vertebrates, cohesins first bind to chromosomes as cells exit mitosis, but the mechanism is unknown3, 11, 12. Concurrent with cohesin binding, pre-replication complexes (pre-RCs) are assembled at origins of DNA replication through the sequential loading of the initiation factors ORC, Cdc6, Cdt1 and MCM2-7 (the 'licensing' reaction)13. In S phase, the protein kinase Cdk2 activates pre-RCs, causing origin unwinding and DNA replication. Here, we use Xenopus egg extracts to show that the recruitment of cohesins to chromosomes requires fully licensed chromatin and is dependent on ORC, Cdc6, Cdt1 and MCM2-7, but is independent of Cdk2. We further show that Xenopus Scc2 is required for cohesin loading and that binding of XScc2 to chromatin is MCM2-7 dependent. Our results define a novel pre-RC-dependent pathway for cohesin recruitment to chromosomes in a vertebrate model system.
- Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
- Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Correspondence to: Johannes C. Walter1 e-mail: johannes_walter@hms.harvard.edu
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