Proliferating human cells hypomorphic for origin recognition complex 2 and pre-replicative complex formation have a defect in p53 activation and Cdk2 kinase activation

Jamie K. Teer, Yuichi Machida, Helene Labit, Olivia Novac, Olivier Hyrien, Kathrin Marheineke, Maria Zannis-Hadjopoulos, Anindya Dutta

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The Origin Recognition Complex (ORC) is a critical component of replication initiation. We have previously reported generation of an Orc2 hypomorph cell line (Δ/-) that expresses very low levels of Orc2 but is viable. We have shown here that Chk2 is phosphorylated, suggesting that DNA damage checkpoint pathways are activated. p53 was inactivated during the derivation of the Orc2 hypomorphic cell lines, accounting for their survival despite active Chk2. These cells also show a defect in the G1 to S-phase transition. Cdk2 kinase activation in G1 is decreased due to decreased Cyclin E levels, preventing progression into S-phase. Molecular combing of bromodeoxyuridine-labeled DNA revealed that once the Orc2 hypomorphic cells enter S-phase, fork density and fork progression are approximately comparable with wild type cells. Therefore, the low level of Orc2 hinders normal cell cycle progression by delaying the activation of G1 cyclin-dependent kinases. The results suggest that hypomorphic mutations in initiation factor genes may be particularly deleterious in cancers with mutant p53 or increased activity of Cyclin E/Cdk2.

Original languageEnglish (US)
Pages (from-to)6253-6260
Number of pages8
JournalJournal of Biological Chemistry
Volume281
Issue number10
DOIs
StatePublished - Mar 10 2006
Externally publishedYes

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Origin Recognition Complex
S Phase
Cyclin E
Phosphotransferases
Chemical activation
Cells
Defects
Cell Line
Peptide Initiation Factors
Cyclin-Dependent Kinases
DNA
Phase Transition
Bromodeoxyuridine
DNA Damage
Cell Cycle
Genes
Phase transitions
Mutation
Survival
Neoplasms

ASJC Scopus subject areas

  • Biochemistry

Cite this

Proliferating human cells hypomorphic for origin recognition complex 2 and pre-replicative complex formation have a defect in p53 activation and Cdk2 kinase activation. / Teer, Jamie K.; Machida, Yuichi; Labit, Helene; Novac, Olivia; Hyrien, Olivier; Marheineke, Kathrin; Zannis-Hadjopoulos, Maria; Dutta, Anindya.

In: Journal of Biological Chemistry, Vol. 281, No. 10, 10.03.2006, p. 6253-6260.

Research output: Contribution to journalArticle

Teer, Jamie K. ; Machida, Yuichi ; Labit, Helene ; Novac, Olivia ; Hyrien, Olivier ; Marheineke, Kathrin ; Zannis-Hadjopoulos, Maria ; Dutta, Anindya. / Proliferating human cells hypomorphic for origin recognition complex 2 and pre-replicative complex formation have a defect in p53 activation and Cdk2 kinase activation. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 10. pp. 6253-6260.
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