title: HJURP uses distinct CENP-A surfaces to recognize and to stabilize CENP-A/histone H4 for centromere assembly.
abstract: Centromeres are defined by the presence of chromatin containing the histone H3 variant, CENP-A, whose assembly into nucleosomes requires the chromatin assembly factor HJURP. We find that whereas surface-exposed residues in the CENP-A targeting domain (CATD) are the primary sequence determinants for HJURP recognition, buried CATD residues that generate rigidity with H4 are also required for efficient incorporation into centromeres. HJURP contact points adjacent to the CATD on the CENP-A surface are not used for binding specificity but rather to transmit stability broadly throughout the histone fold domains of both CENP-A and H4. Furthermore, an intact CENP-A/CENP-A interface is a requirement for stable chromatin incorporation immediately upon HJURP-mediated assembly. These data offer insight into the mechanism by which HJURP discriminates CENP-A from bulk histone complexes and chaperones CENP-A/H4 for a substantial portion of the cell cycle prior to mediating chromatin assembly at the centromere.
authors.affiliation: Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Emily A
authors.initials: EA
authors.lastname: Bassett
authors.forename: Jamie
authors.initials: J
authors.lastname: DeNizio
authors.forename: Meghan C
authors.initials: MC
authors.lastname: Barnhart-Dailey
authors.forename: Tanya
authors.initials: T
authors.lastname: Panchenko
authors.forename: Nikolina
authors.initials: N
authors.lastname: Sekulic
authors.forename: Danielle J
authors.initials: DJ
authors.lastname: Rogers
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Foltz
authors.forename: Ben E
authors.initials: BE
authors.lastname: Black
mesh_terms: D000595:Amino Acid Sequence; D001324:Autoantigens / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D002453:Cell Cycle; D002503:Centromere / Q000502:physiology*; D000076247:Centromere Protein A; D042002:Chromatin Assembly and Disassembly; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000502:physiology; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C524371:HJURP protein, human; D006657:Histones; D009707:Nucleosomes; D011994:Recombinant Proteins