title: Replication of alpha-satellite DNA arrays in endogenous human centromeric regions and in human artificial chromosome.
abstract: In human chromosomes, centromeric regions comprise megabase-size arrays of 171 bp alpha-satellite DNA monomers. The large distances spanned by these arrays preclude their replication from external sites and imply that the repetitive monomers contain replication origins. However, replication within these arrays has not previously been profiled and the role of alpha-satellite DNA in initiation of DNA replication has not yet been demonstrated. Here, replication of alpha-satellite DNA in endogenous human centromeric regions and in de novo formed Human Artificial Chromosome (HAC) was analyzed. We showed that alpha-satellite monomers could function as origins of DNA replication and that replication of alphoid arrays organized into centrochromatin occurred earlier than those organized into heterochromatin. The distribution of inter-origin distances within centromeric alphoid arrays was comparable to the distribution of inter-origin distances on randomly selected non-centromeric chromosomal regions. Depletion of CENP-B, a kinetochore protein that binds directly to a 17 bp CENP-B box motif common to alpha-satellite DNA, resulted in enrichment of alpha-satellite sequences for proteins of the ORC complex, suggesting that CENP-B may have a role in regulating the replication of centromeric regions. Mapping of replication initiation sites in the HAC revealed that replication preferentially initiated in transcriptionally active regions.
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Indri
authors.initials: I
authors.lastname: Erliandri
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Haiqing
authors.initials: H
authors.lastname: Fu
authors.affiliation: Laboratory of Cell Engineering, Department of Frontier Research, Kazusa DNA Research Institute, Kisarazu, Chiba, 292-0818, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Jung-Hyun
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Center for Biomolecular Science and Engineering, University of California, Santa Cruz, CA, 95064, USA.
authors.forename: Karen H
authors.initials: KH
authors.lastname: Miga
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Liskovykh
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3JR, UK.
authors.forename: William C
authors.initials: WC
authors.lastname: Earnshaw
authors.affiliation: Laboratory of Cell Engineering, Department of Frontier Research, Kazusa DNA Research Institute, Kisarazu, Chiba, 292-0818, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masumoto
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA kouprinn@mail.nih.gov.
authors.forename: Natalay
authors.initials: N
authors.lastname: Kouprina
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Mirit I
authors.initials: MI
authors.lastname: Aladjem
authors.affiliation: Developmental Therapeutics Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Larionov
mesh_terms: D002460:Cell Line; D045744:Cell Line, Tumor; D002503:Centromere; D051936:Centromere Protein B / Q000502:physiology; D022222:Chromosomes, Artificial, Human; D004261:DNA Replication; D042522:DNA Replication Timing; D004276:DNA, Satellite / Q000096:biosynthesis*; D006801:Humans; D007202:Indicators and Reagents; D018741:Replication Origin
chemical_list: D051936:Centromere Protein B; D004276:DNA, Satellite; D007202:Indicators and Reagents