title: HJURP is a CENP-A chromatin assembly factor sufficient to form a functional de novo kinetochore.
abstract: Centromeres of higher eukaryotes are epigenetically marked by the centromere-specific CENP-A nucleosome. New CENP-A recruitment requires the CENP-A histone chaperone HJURP. In this paper, we show that a LacI (Lac repressor) fusion of HJURP drove the stable recruitment of CENP-A to a LacO (Lac operon) array at a noncentromeric locus. Ectopically targeted CENP-A chromatin at the LacO array was sufficient to direct the assembly of a functional centromere as indicated by the recruitment of the constitutive centromere-associated network proteins, the microtubule-binding protein NDC80, and the formation of stable kinetochore-microtubule attachments. An amino-terminal fragment of HJURP was able to assemble CENP-A nucleosomes in vitro, demonstrating that HJURP is a chromatin assembly factor. Furthermore, HJURP recruitment to endogenous centromeres required the Mis18 complex. Together, these data suggest that the role of the Mis18 complex in CENP-A deposition is to recruit HJURP and that the CENP-A nucleosome assembly activity of HJURP is responsible for centromeric chromatin assembly to maintain the epigenetic mark.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Virginia Medical School, Charlottesville, VA 22908, USA.
authors.forename: Meghan C
authors.initials: MC
authors.lastname: Barnhart
authors.forename: P Henning J L
authors.initials: PH
authors.lastname: Kuich
authors.forename: Madison E
authors.initials: ME
authors.lastname: Stellfox
authors.forename: Jared A
authors.initials: JA
authors.lastname: Ward
authors.forename: Emily A
authors.initials: EA
authors.lastname: Bassett
authors.forename: Ben E
authors.initials: BE
authors.lastname: Black
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Foltz
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D002478:Cells, Cultured; D000076247:Centromere Protein A; D042002:Chromatin Assembly and Disassembly; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; C000619701:Cenpa protein, mouse; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C524371:HJURP protein, human