BACKGROUND: Sister chromatids are held together by the ring-shaped cohesin complex, which is loaded onto chromosomes before DNA replication. Cohesion between sister chromosomes is established during DNA replication, and it requires acetylation of the Smc3 subunit of cohesin by evolutionally conserved cohesin acetyltransferases (CoATs). However, how CoATs are recruited to chromatin and how cohesin acetylation is regulated remain unclear. RESULTS: We found that cohesin acetylation requires pre-RC-dependent chromatin loading of cohesin, but surprisingly, it is independent of DNA synthesis in Xenopus egg extracts. Immunodepletion experiments revealed that XEco2 is the CoAT responsible for Smc3 acetylation and sister chromatid cohesion. Recruitment of XEco2 onto chromatin was dependent on pre-RC assembly but was independent of cohesin loading and DNA synthesis. Two short N-terminal motifs, PBM-A and PBM-B, which are conserved among vertebrate Esco2/XEco2 homologs, were collectively essential for pre-RC-dependent chromatin association of XEco2, cohesin acetylation, and subsequent sister chromatid cohesion. The conserved PCNA-interacting protein box in XEco2 was largely dispensable for Smc3 acetylation but was partially required for cohesion. Interaction of acetylated cohesin with DNA was stabilized against salt-wash treatments after DNA replication. CONCLUSIONS: 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.
BACKGROUND: Sister chromatids are held together by the ring-shaped cohesin complex, which is loaded onto chromosomes before DNA replication. Cohesion between sister chromosomes is established during DNA replication, and it requires acetylation of the Smc3 subunit of cohesin by evolutionally conserved cohesin acetyltransferases (CoATs). However, how CoATs are recruited to chromatin and how cohesin acetylation is regulated remain unclear. RESULTS: We found that cohesin acetylation requires pre-RC-dependent chromatin loading of cohesin, but surprisingly, it is independent of DNA synthesis in Xenopus egg extracts. Immunodepletion experiments revealed that XEco2 is the CoAT responsible for Smc3 acetylation and sister chromatid cohesion. Recruitment of XEco2 onto chromatin was dependent on pre-RC assembly but was independent of cohesin loading and DNA synthesis. Two short N-terminal motifs, PBM-A and PBM-B, which are conserved among vertebrate Esco2/XEco2 homologs, were collectively essential for pre-RC-dependent chromatin association of XEco2, cohesin acetylation, and subsequent sister chromatid cohesion. The conserved PCNA-interacting protein box in XEco2 was largely dispensable for Smc3 acetylation but was partially required for cohesion. Interaction of acetylated cohesin with DNA was stabilized against salt-wash treatments after DNA replication. CONCLUSIONS: 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.
Authors: Miroslav P Ivanov; Rene Ladurner; Ina Poser; Rebecca Beveridge; Evelyn Rampler; Otto Hudecz; Maria Novatchkova; Jean-Karim Hériché; Gordana Wutz; Petra van der Lelij; Emanuel Kreidl; James Ra Hutchins; Heinz Axelsson-Ekker; Jan Ellenberg; Anthony A Hyman; Karl Mechtler; Jan-Michael Peters Journal: EMBO J Date: 2018-06-21 Impact factor: 11.598
Authors: Vladimir P Bermudez; Andrea Farina; Torahiko L Higashi; Fang Du; Inger Tappin; Tatsuro S Takahashi; Jerard Hurwitz Journal: Proc Natl Acad Sci U S A Date: 2012-05-24 Impact factor: 11.205
Authors: Reem M Alomer; Eulália M L da Silva; Jingrong Chen; Katarzyna M Piekarz; Katherine McDonald; Courtney G Sansam; Christopher L Sansam; Susannah Rankin Journal: Proc Natl Acad Sci U S A Date: 2017-08-28 Impact factor: 11.205
Authors: Dawn Bender; Eulália Maria Lima Da Silva; Jingrong Chen; Annelise Poss; Lauren Gawey; Zane Rulon; Susannah Rankin Journal: Proc Natl Acad Sci U S A Date: 2019-12-26 Impact factor: 11.205