XRCC1 and DNA polymerase beta in cellular protection against cytotoxic DNA single-strand breaks.
Single-strand breaks (SSBs) can occur in cells either directly, or indirectly following initiation of base excision repair (BER). SSBs generally have blocked termini lacking the conventional 5'-phosphate and 3'-hydroxyl groups and require further processing prior to DNA synthesis and ligation. XRCC1 is devoid of any known enzymatic activity, but it can physically interact with other proteins involved in all stages of the overlapping SSB repair and BER pathways, including those that conduct the rate-limiting end-tailoring, and in many cases can stimulate their enzymatic activities. XRCC1(-/-) mouse fibroblasts are most hypersensitive to agents that produce DNA lesions repaired by monofunctional glycosylase-initiated BER and that result in formation of indirect SSBs. A requirement for the deoxyribose phosphate lyase activity of DNA polymerase beta (pol beta) is specific to this pathway, whereas pol beta is implicated in gap-filling during repair of many types of SSBs. Elevated levels of strand breaks, and diminished repair, have been demonstrated in MMS-treated XRCC1(-/-), and to a lesser extent in pol beta(-/-) cell lines, compared with wild-type cells. Thus a strong correlation is observed between cellular sensitivity to MMS and the ability of cells to repair MMS-induced damage. Exposure of wild-type and pol beta(-/-) cells to an inhibitor of PARP activity dramatically potentiates MMS-induced cytotoxicity. XRCC1(-/-) cells are also sensitized by PARP inhibition demonstrating that PARP-mediated poly(ADP-ribosyl)ation plays a role in modulation of cytotoxicity beyond recruitment of XRCC1 to sites of DNA damage.
Horton
Julie K
JK
Laboratory of Structural Biology, NIEHS, National Institutes of Health, Research Triangle Park, NC 27709, USA.
Watson
Mary
M
Stefanick
Donna F
DF
Shaughnessy
Daniel T
DT
Taylor
Jack A
JA
Wilson
Samuel H
SH
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D019610:Cytoprotection / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D053904:DNA Breaks, Single-Stranded; D019951:DNA Polymerase beta / Q000235:genetics / Q000502:physiology*; D049914:DNA Repair-Deficiency Disorders / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008795:Metronidazole / Q000031:analogs & derivatives / Q000633:toxicity; D009153:Mutagens / Q000633:toxicity; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D011839:Radiation, Ionizing; D000076105:X-ray Repair Cross Complementing Protein 1
D004268:DNA-Binding Proteins; D009153:Mutagens; D000076105:X-ray Repair Cross Complementing Protein 1; C000619371:XRCC1 protein, human; C000619352:Xrcc1 protein, mouse; C051964:metronidazole monosuccinate; D008795:Metronidazole; D019951:DNA Polymerase beta