CDC-48/p97 coordinates CDT-1 degradation with GINS chromatin dissociation to ensure faithful DNA replication

Mol Cell. 2011 Oct 7;44(1):85-96. doi: 10.1016/j.molcel.2011.08.028.

Abstract

Faithful transmission of genomic information requires tight spatiotemporal regulation of DNA replication factors. In the licensing step of DNA replication, CDT-1 is loaded onto chromatin to subsequently promote the recruitment of additional replication factors, including CDC-45 and GINS. During the elongation step, the CDC-45/GINS complex moves with the replication fork; however, it is largely unknown how its chromatin association is regulated. Here, we show that the chaperone-like ATPase CDC-48/p97 coordinates degradation of CDT-1 with release of the CDC-45/GINS complex. C. elegans embryos lacking CDC-48 or its cofactors UFD-1/NPL-4 accumulate CDT-1 on mitotic chromatin, indicating a critical role of CDC-48 in CDT-1 turnover. Strikingly, CDC-48(UFD-1/NPL-4)-deficient embryos show persistent chromatin association of CDC-45/GINS, which is a consequence of CDT-1 stabilization. Moreover, our data confirmed a similar regulation in Xenopus egg extracts, emphasizing a conserved coordination of licensing and elongation events during eukaryotic DNA replication by CDC-48/p97.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism*
  • Animals
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Chromatin / metabolism*
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA Replication*
  • Ligases / metabolism*
  • Male
  • Mitosis
  • RNA Interference
  • Spermatozoa / metabolism
  • Two-Hybrid System Techniques
  • Ubiquitin / chemistry
  • Ubiquitin / metabolism
  • Valosin Containing Protein
  • Xenopus Proteins / metabolism*
  • Xenopus laevis

Substances

  • Caenorhabditis elegans Proteins
  • Cdc45 protein, Xenopus
  • Cdt-1 protein, C elegans
  • Cell Cycle Proteins
  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • Ubiquitin
  • Xenopus Proteins
  • evl-18 protein, C elegans
  • Adenosine Triphosphatases
  • Valosin Containing Protein
  • Ligases