A CAF-1-PCNA-mediated chromatin assembly pathway triggered by sensing DNA damage

Mol Cell Biol. 2000 Feb;20(4):1206-18. doi: 10.1128/MCB.20.4.1206-1218.2000.

Abstract

Sensing DNA damage is crucial for the maintenance of genomic integrity and cell cycle progression. The participation of chromatin in these events is becoming of increasing interest. We show that the presence of single-strand breaks and gaps, formed either directly or during DNA damage processing, can trigger the propagation of nucleosomal arrays. This nucleosome assembly pathway involves the histone chaperone chromatin assembly factor 1 (CAF-1). The largest subunit (p150) of this factor interacts directly with proliferating cell nuclear antigen (PCNA), and critical regions for this interaction on both proteins have been mapped. To isolate proteins specifically recruited during DNA repair, damaged DNA linked to magnetic beads was used. The binding of both PCNA and CAF-1 to this damaged DNA was dependent on the number of DNA lesions and required ATP. Chromatin assembly linked to the repair of single-strand breaks was disrupted by depletion of PCNA from a cell-free system. This defect was rescued by complementation with recombinant PCNA, arguing for role of PCNA in mediating chromatin assembly linked to DNA repair. We discuss the importance of the PCNA-CAF-1 interaction in the context of DNA damage processing and checkpoint control.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites / genetics
  • Cell-Free System
  • Chromatin / genetics
  • Chromatin / metabolism*
  • Chromatin Assembly Factor-1
  • Chromosomal Proteins, Non-Histone*
  • DNA / biosynthesis
  • DNA Damage*
  • DNA Primers / genetics
  • DNA Repair
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Drosophila
  • HeLa Cells
  • Humans
  • In Vitro Techniques
  • Models, Biological
  • Models, Molecular
  • Nucleosomes / metabolism
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism*
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transcription Factors

Substances

  • CHAF1B protein, human
  • CNOT8 protein, human
  • Chromatin
  • Chromatin Assembly Factor-1
  • Chromosomal Proteins, Non-Histone
  • DNA Primers
  • DNA-Binding Proteins
  • Nucleosomes
  • Proliferating Cell Nuclear Antigen
  • Recombinant Proteins
  • Transcription Factors
  • DNA