Human Orc2 localizes to centrosomes, centromeres and heterochromatin during chromosome inheritance

Supriya G. Prasanth, Kannanganattu V. Prasanth, Khalid Siddiqui, David L. Spector, Bruce Stillman

Research output: Contribution to journalArticlepeer-review

Abstract

The initiation of DNA replication in S phase requires the prior assembly of an origin recognition complex (ORC)-dependent pre-replicative complex on chromatin during G1 phase of the cell division cycle. In human cells, the Orc2 subunit localized to the nucleus as expected, but it also localized to centrosomes throughout the entire cell cycle. Furthermore, Orc2 was tightly bound to heterochromatin and heterochromatin protein 1α (HP1α) and HP1β in G1 and early S phase, but during late S, G2 and M phases tight chromatin association was restricted to centromeres. Depletion of Orc2 by siRNA caused multiple phenotypes. A population of cells showed an S-phase defect with little proliferating cell nuclear antigen (PCNA) on chromatin, although MCM proteins remained. Orc2 depletion also disrupted HP1 localization, but not histone-H3-lysine-9 methylation at prominent heterochromatic foci. Another subset of Orc2-depleted cells containing replicated DNA arrested with abnormally condensed chromosomes, failed chromosome congression and multiple centrosomes. These results implicate Orc2 protein in chromosome du-plication, chromosome structure and centrosome copy number control, suggesting that it coordinates all stages of the chromosome inheritance cycle.

Original languageEnglish (US)
Pages (from-to)2651-2663
Number of pages13
JournalEMBO Journal
Volume23
Issue number13
DOIs
StatePublished - Jul 7 2004
Externally publishedYes

Keywords

  • Centromeres
  • Centrosomes
  • DNA replication
  • Heterochromatin
  • Origin recognition complex

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Fingerprint

Dive into the research topics of 'Human Orc2 localizes to centrosomes, centromeres and heterochromatin during chromosome inheritance'. Together they form a unique fingerprint.

Cite this