XRCC1 is phosphorylated by DNA-dependent protein kinase in response to DNA damage

The two BRCT domains (BRCT1 and BRCT2) of XRCC1 mediate a network of protein–protein interactions with several key factors of the DNA single-strand breaks (SSBs) and base damage repair pathways. BRCT1 is required for the immediate poly(ADP–ribose)-dependent recruitment of XRCC1 to DNA breaks and is...

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Bibliographic Details
Published in:Nucleic acids research 2006-01, Vol.34 (1), p.32-41
Main Authors: Lévy, Nicolas, Martz, Adeline, Bresson, Anne, Spenlehauer, Catherine, de Murcia, Gilbert, Ménissier-de Murcia, Josiane
Format: Article
Language:English
Subjects:
Animals
Binding Sites
Cricetinae
Dimerization
DNA Damage
DNA Repair
DNA-Activated Protein Kinase - metabolism
DNA-Binding Proteins - chemistry
DNA-Binding Proteins - metabolism
HeLa Cells
Humans
Mass Spectrometry
Phosphorylation
Protein Structure, Tertiary
Radiation, Ionizing
Serine - metabolism
X-ray Repair Cross Complementing Protein 1
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Summary:The two BRCT domains (BRCT1 and BRCT2) of XRCC1 mediate a network of protein–protein interactions with several key factors of the DNA single-strand breaks (SSBs) and base damage repair pathways. BRCT1 is required for the immediate poly(ADP–ribose)-dependent recruitment of XRCC1 to DNA breaks and is essential for survival after DNA damage. To better understand the biological role of XRCC1 in the processing of DNA ends, a search for the BRCT1 domain-associated proteins was performed by mass spectrometry of GST-BRCT1 pulled-down proteins from HeLa cell extracts. Here, we report that the double-strand break (DSB) repair heterotrimeric complex DNA-PK interacts with the BRCT1 domain of XRCC1 and phosphorylates this domain at serine 371 after ionizing irradiation. This caused XRCC1 dimer dissociation. The XRCC1 R399Q variant allele did not affect this phosphorylation. We also show that XRCC1 strongly stimulates the phosphorylation of p53-Ser15 by DNA-PK. The pseudo phosphorylated S371D mutant was a much weaker stimulator of DNA-PK activity whereas the non-phosphorylable mutant S371L endowed with a DNA-PK stimulating capacity failed to fully rescue the DSB repair defect of XRCC1-deficient EM9 rodent cells. The functional association between XRCC1 and DNA-PK in response to IR provides the first evidence for their involvement in a common DSB repair pathway.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkj409