Disruption of nuclear lamin organization blocks the elongation phase of DNA replication

Robert D. Moir, Timothy P. Spann, Harald Herrmann, Robert D. Goldman*

*Corresponding author for this work

Research output: Contribution to journalArticle

167 Scopus citations

Abstract

The role of nuclear lamins in DNA replication is unclear. To address this, nuclei were assembled in Xenopus extracts containing AraC, a reversible inhibitor that blocks near the onset of the elongation phase of replication. Dominant-negative lamin mutants lacking their NH2-terminal domains were added to assembled nuclei to disrupt lamin organization. This prevented the resumption of DNA replication after the release of the AraC block. This inhibition of replication was not due to gross disruption of nuclear envelope structure and function. The organization of initiation factors was not altered by lamin disruption, and nuclei resumed replication when transferred to extracts treated with CIP, an inhibitor of the cyclin-dependent kinase (cdk) 2-dependent step of initiation. This suggests that alteration of lamin organization does not affect the initiation phase of DNA replication. Instead, we find that disruption of lamin organization inhibited chain elongation in a dose-dependent fashion. Furthermore, the established organization of two elongation factors, proliferating cell nuclear antigen, and replication factor complex, was disrupted by ΔNLA. These findings demonstrate that lamin organization must be maintained in nuclei for the elongation phase of DNA replication to proceed.

Original languageEnglish (US)
Pages (from-to)1179-1191
Number of pages13
JournalJournal of Cell Biology
Volume149
Issue number6
DOIs
StatePublished - Jun 12 2000

Keywords

  • DNA synthesis
  • Intermediate filaments
  • Lamin mutant
  • Lamina
  • Nuclear organization

ASJC Scopus subject areas

  • Cell Biology

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