Discovery - University of Dundee - Online Publications

Library & Learning Centre

The dynamics of replication licensing in live Caenorhabditis elegans embryos

The dynamics of replication licensing in live Caenorhabditis elegans embryos

Research output: Contribution to journalArticle

View graph of relations

Authors

  • Remi Sonneville
  • Matthieu Querenet
  • Ashley Craig
  • Anton Gartner (Lead / Corresponding author)
  • J. Julian Blow (Lead / Corresponding author)

Research units

Info

Original languageEnglish
Pages233-246
Number of pages14
JournalJournal of Cell Biology
Journal publication date23 Jan 2012
Journal number2
Volume196
DOIs
StatePublished

Abstract

Accurate DNA replication requires proper regulation of replication licensing, which entails loading MCM-2-7 onto replication origins. In this paper, we provide the first comprehensive view of replication licensing in vivo, using video microscopy of Caenorhabditis elegans embryos. As expected, MCM-2-7 loading in late M phase depended on the prereplicative complex (pre-RC) proteins: origin recognition complex (ORC), CDC-6, and CDT-1. However, many features we observed have not been described before: GFP-ORC-1 bound chromatin independently of ORC-2-5, and CDC-6 bound chromatin independently of ORC, whereas CDT-1 and MCM-2-7 DNA binding was interdependent. MCM-3 chromatin loading was irreversible, but CDC-6 and ORC turned over rapidly, consistent with ORC/CDC-6 loading multiple MCM-2-7 complexes. MCM-2-7 chromatin loading further reduced ORC and CDC-6 DNA binding. This dynamic behavior creates a feedback loop allowing ORC/CDC-6 to repeatedly load MCM-2-7 and distribute licensed origins along chromosomal DNA. During S phase, ORC and CDC-6 were excluded from nuclei, and DNA was overreplicated in export-defective cells. Thus, nucleocytoplasmic compartmentalization of licensing factors ensures that DNA replication occurs only once.

Download statistics

No data available

Documents

Documents

DOI

Library & Learning Centre

Contact | Accessibility | Policy