Chromatin structure imposed by condensin II at centromeres enables xHJURP-mediated incorporation of CENP-A

Almouzni, Geneviève

Arnaoutov, Alexei

Bernad, Rafael

Boyarchuk, Ekaterina

Dasso, Mary

Losada, Ana

Ray-Gallet, Dominique

Rivera, Teresa

Rodríguez-Corsino, Miriam

Sánchez, Patricia

Vassias, Isabelle

English

PubMed

Centromeric protein A (CENP-A) is the epigenetic mark of centromeres. CENP-A replenishment is necessary in each cell cycle to compensate for the dilution associated to DNA replication, but how this is achieved mechanistically is largely unknown. We have developed an assay using Xenopus egg extracts that can recapitulate the spatial and temporal specificity of CENP-A deposition observed in human cells, providing us with a robust in vitro system amenable to molecular dissection. Here we show that this deposition depends on Xenopus Holliday junction-recognizing protein (xHJURP), a member of the HJURP/Scm3 family recently identified in yeast and human cells, further supporting the essential role of these chaperones in CENP-A loading. Despite little sequence homology, human HJURP can substitute for xHJURP. We also report that condensin II, but not condensin I, is required for CENP-A assembly and contributes to retention of centromeric CENP-A nucleosomes both in mitosis and interphase. We propose that the chromatin structure imposed by condensin II at centromeres enables CENP-A incorporation initiated by xHJURP.This research has been supported by the Spanish Ministry of Science and Innovation (grants BFU2007-66627 and CSD-Inesgen to A. Losada, a Juan de la Cierva contract for R. Bernad, and FPI fellowships to P. Sanchez and T. Rivera); the European Commission Epigenome Network of Excellence (LSHG-CT-2004-503433) to G. Almouzni and A. Losada; MIRG-31126 to A. Losada; and EMBO fellowship ALTF 77-2007 for R. Bernad. Further support for the group of G. Almouzni comes from La Ligue Nationale contre le Cancer (Equipe labelisee Ligue 2010); PIC Programs ("Retinoblastome" and "Replication, Instabilite chromosomique et cancer"); European Commission ITN FP7-PEOPLE-2007 "Image DDR" and FP7-PEOPLE-2008 "Nucleosome 4D"; ACI-2007-Canceropole IdF "Breast cancer and Epigenetics"; ANR "FaRC" PCV06_142302 and ANR "ECenS" ANR-09-BLAN-0257-01; and INCa "GepiG".S

REPISALUD (Instituto de Salud Carlos III)

The Journal of Cell Biology

Xenopus HJURP and condensin II are required for CENP-A assembly.

Centromeric protein A (CENP-A) is the epigenetic mark of centromeres. CENP-A replenishment is necessary in each cell cycle to compensate for the dilution associated to DNA replication, but how this is achieved mechanistically is largely unknown. We have developed an assay using Xenopus egg extracts that can recapitulate the spatial and temporal specificity of CENP-A deposition observed in human cells, providing us with a robust in vitro system amenable to molecular dissection. Here we show that this deposition depends on Xenopus Holliday junction–recognizing protein (xHJURP), a member of the HJURP/Scm3 family recently identified in yeast and human cells, further supporting the essential role of these chaperones in CENP-A loading. Despite little sequence homology, human HJURP can substitute for xHJURP. We also report that condensin II, but not condensin I, is required for CENP-A assembly and contributes to retention of centromeric CENP-A nucleosomes both in mitosis and interphase. We propose that the chromatin structure imposed by condensin II at centromeres enables CENP-A incorporation initiated by xHJURP.</jats:p

Rafael Bernad

Patricia Sánchez

Teresa Rivera

Miriam Rodríguez-Corsino

Ekaterina Boyarchuk

Isabelle Vassias

Dominique Ray-Gallet

Alexei Arnaoutov

Mary Dasso

Geneviève Almouzni

Ana Losada

Crossref

Xenopus HJURP and condensin II are required for CENP-A assembly

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http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3051821

Xenopus HJURP and condensin II are required for CENP-A assembly

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REPISALUD (Instituto de Salud Carlos III)

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