title: The WD40 Domain of HIRA Is Essential for RI-nucleosome Assembly in Xenopus Egg Extracts.
abstract: Histone chaperones are a group of histone-binding proteins that facilitate the assembly of nucleosomes, the fundamental structural units of chromatin in eukaryotes. In nucleosome assembly, deposition of a histone H3-H4 tetramer onto DNA is the first and critical step, which is mediated by the histone chaperones HIRA and CAF-1. HIRA and CAF-1 are reportedly involved in DNA replication independent (RI) and replication coupled nucleosome assembly, respectively. However, the mechanisms by which they mediate histone deposition remain unclear. In this study, we focused on the mechanism by which HIRA induces RI-nucleosome assembly. We looked for HIRA domains that are required for nucleosome assembly and its localization to chromatin. We used cell-free extracts from Xenopus eggs that carry out RI-nucleosome assembly of plasmid DNA. We confirmed that HIRA formed stable complexes with Asf1, another histone H3-H4 chaperone, and the HIRA-Asf1 complex was solely responsible for RI-nucleosome assembly in egg extracts. We further demonstrated that the HIRA N-terminus containing the WD40 domain, which comprises seven WD40 repeats, and the B domain, to which Asf1 binds, were essential for RI-nucleosome assembly; the three WD40 repeats from the N-terminus were especially critical. Using egg extracts that reproduce nuclear formation accompanying the duplication of chromatin, we also demonstrated that the Hir domain was indispensable for the binding of HIRA to chromatin. Thus, the WD40 and B domains are the core elements for inducing RI-nucleosome assembly. Hir domain regulates the binding to chromatin. Based on these findings, similarities and differences between HIRA and CAF-1 are discussed.
authors.affiliation: Group of Developmental Cell Biology, Graduate School of Science, Nagoya University.
authors.forename: Ruibin
authors.initials: R
authors.lastname: Zhu
authors.forename: Mari
authors.initials: M
authors.lastname: Iwabuchi
authors.forename: Keita
authors.initials: K
authors.lastname: Ohsumi
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000737:chemistry; D002457:Cell Extracts; D004261:DNA Replication; D056488:Histone Chaperones / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D009707:Nucleosomes / Q000378:metabolism*; D009865:Oocytes; D014157:Transcription Factors / Q000737:chemistry; D000071678:WD40 Repeats; D014982:Xenopus laevis
chemical_list: D018797:Cell Cycle Proteins; D002457:Cell Extracts; C086057:HIRA protein, human; D056488:Histone Chaperones; D009707:Nucleosomes; D014157:Transcription Factors