title: Centromere identity maintained by nucleosomes assembled with histone H3 containing the CENP-A targeting domain.
abstract: Active centromeres are marked by nucleosomes assembled with CENP-A, a centromere-specific histone H3 variant. The CENP-A centromere targeting domain (CATD), comprised of loop 1 and the alpha2 helix within the histone fold, is sufficient to target histone H3 to centromeres and to generate the same conformational rigidity to the initial subnucleosomal heterotetramer with histone H4 as does CENP-A. We now show in human cells and in yeast that depletion of CENP-A is lethal, but recruitment of normal levels of kinetochore proteins, centromere-generated mitotic checkpoint signaling, chromosome segregation, and viability can be rescued by histone H3 carrying the CATD. These data offer direct support for centromere identity maintained by a unique nucleosome that serves to distinguish the centromere from the rest of the chromosome.
authors.affiliation: Ludwig Institute for Cancer Research, University of California, San Diego, La Jolla, CA 92093, USA. blackbe@mail.med.upenn.edu
authors.forename: Ben E
authors.initials: BE
authors.lastname: Black
authors.forename: Lars E T
authors.initials: LE
authors.lastname: Jansen
authors.forename: Paul S
authors.initials: PS
authors.lastname: Maddox
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Foltz
authors.forename: Arshad B
authors.initials: AB
authors.lastname: Desai
authors.forename: Jagesh V
authors.initials: JV
authors.lastname: Shah
authors.forename: Don W
authors.initials: DW
authors.lastname: Cleveland
mesh_terms: D001324:Autoantigens / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002503:Centromere / Q000378:metabolism*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008938:Mitosis; D008954:Models, Biological; D009707:Nucleosomes / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D001324:Autoantigens; D001426:Bacterial Proteins; C000619700:CENPA protein, human; C092746:CSE4 protein, S cerevisiae; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; D008164:Luminescent Proteins; D009707:Nucleosomes; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C054562:yellow fluorescent protein, Bacteria