XPA-mediated regulation of global nucleotide excision repair by ATR Is p53-dependent and occurs primarily in S-phase

PLoS One. 2011;6(12):e28326. doi: 10.1371/journal.pone.0028326. Epub 2011 Dec 12.

Abstract

Cell cycle checkpoints play an important role in regulation of DNA repair pathways. However, how the regulation occurs throughout the cell cycle remains largely unknown. Here we demonstrate that nucleotide excision repair (NER) is regulated by the ATR/p53 checkpoint via modulation of XPA nuclear import and that this regulation occurs in a cell cycle-dependent manner. We show that depletion of p53 abrogated the UV-induced nuclear translocation of XPA, while silencing of Chk1 or MAPKAP Kinase-2 (MK2) had no effect. Inhibition of p53 transcriptional activities and silencing of p53-Ser15 phosphorylation also reduced the damage-induced XPA nuclear import. Furthermore, in G1-phase cells the majority of XPA remained in the cytoplasm even after UV treatment. By contrast, while most of the XPA in S-phase cells was initially located in the cytoplasm before DNA damage, UV irradiation stimulated bulk import of XPA into the nucleus. Interestingly, the majority of XPA molecules always were located in the nucleus in G2-phase cells no matter whether the DNA was damaged or not. Consistently, the UV-induced Ser15 phosphorylation of p53 occurred mainly in S-phase cells, and removal of cyclobutane pyrimidine dimers (CPDs) was much more efficient in S-phase cells than in G1-phase cells. Our results suggest that upon DNA damage in S phase, NER could be regulated by the ATR/p53-dependent checkpoint via modulation of the XPA nuclear import process. In contrast, the nuclear import of XPA in G(1) or G(2) phase appears to be largely independent of DNA damage and p53.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Active Transport, Cell Nucleus / radiation effects
  • Ataxia Telangiectasia Mutated Proteins
  • Cell Cycle Checkpoints / radiation effects
  • Cell Cycle Proteins / metabolism*
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Cell Nucleus / radiation effects
  • Checkpoint Kinase 1
  • DNA Damage
  • DNA Repair* / radiation effects
  • G1 Phase / radiation effects
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Phosphorylation / radiation effects
  • Phosphoserine / metabolism
  • Protein Kinases / metabolism
  • Protein Serine-Threonine Kinases / metabolism*
  • Protein Transport / radiation effects
  • Pyrimidine Dimers / metabolism
  • S Phase* / radiation effects
  • Subcellular Fractions / metabolism
  • Subcellular Fractions / radiation effects
  • Tumor Suppressor Protein p53 / metabolism
  • Ultraviolet Rays
  • Xeroderma Pigmentosum Group A Protein / metabolism*

Substances

  • Cell Cycle Proteins
  • Intracellular Signaling Peptides and Proteins
  • Pyrimidine Dimers
  • Tumor Suppressor Protein p53
  • XPA protein, human
  • Xeroderma Pigmentosum Group A Protein
  • Phosphoserine
  • Protein Kinases
  • MAP-kinase-activated kinase 2
  • ATR protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • CHEK1 protein, human
  • Checkpoint Kinase 1
  • Protein Serine-Threonine Kinases