title: Xenopus HJURP and condensin II are required for CENP-A assembly.
abstract: 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.
authors.affiliation: Molecular Oncology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Bernad
authors.forename: Patricia
authors.initials: P
authors.lastname: Sánchez
authors.forename: Teresa
authors.initials: T
authors.lastname: Rivera
authors.forename: Miriam
authors.initials: M
authors.lastname: Rodríguez-Corsino
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Boyarchuk
authors.forename: Isabelle
authors.initials: I
authors.lastname: Vassias
authors.forename: Dominique
authors.initials: D
authors.lastname: Ray-Gallet
authors.forename: Alexei
authors.initials: A
authors.lastname: Arnaoutov
authors.forename: Mary
authors.initials: M
authors.lastname: Dasso
authors.forename: Geneviève
authors.initials: G
authors.lastname: Almouzni
authors.forename: Ana
authors.initials: A
authors.lastname: Losada
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D002503:Centromere / Q000378:metabolism / Q000648:ultrastructure; D000076247:Centromere Protein A; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D044127:Epigenesis, Genetic; D056488:Histone Chaperones / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007399:Interphase; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014982:Xenopus laevis
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C524371:HJURP protein, human; D056488:Histone Chaperones; C557677:Holliday junction recognizing protein, Xenopus; D046912:Multiprotein Complexes; D029867:Xenopus Proteins; C108312:condensin complexes; D000251:Adenosine Triphosphatases