TY - JOUR
T1 - XRCC1 interactions with multiple DNA glycosylases
T2 - A model for its recruitment to base excision repair
AU - Campalans, Anna
AU - Marsin, Stéphanie
AU - Nakabeppu, Yusaku
AU - O'Connor, Timothy R.
AU - Boiteux, Serge
AU - Radicella, J. Pablo
N1 - Funding Information:
We would like to thank the staff of the Plate-Forme Imagerie et Biologie Cellulaire, CNRS, Gif sur Yvette, for their help with confocal microscopy. We are grateful to Dr. Daisuke Tsuchimoto for his comments on the manuscript and together with Dr. Kumiko Torisu for their confirmation of DNA sequence of Neil 1 and 2 cDNAs. This work was supported by the Association pour la Recherche sur le Cancer (ARC 4688), Electricité de France (EDF) and EC grant FIGH-CT 2002-0027.
PY - 2005/7/12
Y1 - 2005/7/12
N2 - Repair of chemically modified bases in DNA is accomplished through base excision repair (BER). This pathway is initiated by a specific DNA glycosylase that recognizes and excises the altered base to yield an abasic (AP) site. After cleavage of the AP site by APE1, repair proceeds through re-synthesis and ligation steps. In mammalian cells, the XRCC1 protein, essential for the maintenance of genomic stability, is involved in both base excision and single-strand break repair. XRCC1 participates in the first step of BER by interacting with the human DNA glycosylases hOGG1 and NEIL1. To analyze the possibility of a general mechanism involving the interaction of XRCC1 with DNA glycosylases we used XRCC1 to pull-down DNA glycosylases activities from human cell extracts. XRCC1 co-purifies with DNA glycosylase activities capable of excising hypoxanthine and dihydrothymine, in addition to 8-oxoguanine, but not uracil. Biochemical analyses with the purified proteins confirmed the interactions between XRCC1 and MPG, hNTH1 or hNEIL2. Furthermore, XRCC1 stimulates the activities of these enzymes. In vivo localization studies show that after genotoxic treatments these DNA glycosylases can be found associated with XRCC1 foci. Our results support a BER model in which XRCC1 is recruited to the repair of alkylated or oxidized bases by the enzyme recognizing the lesion. XRCC1 would then coordinate the subsequent enzymatic steps and modulate the activities of all the proteins involved.
AB - Repair of chemically modified bases in DNA is accomplished through base excision repair (BER). This pathway is initiated by a specific DNA glycosylase that recognizes and excises the altered base to yield an abasic (AP) site. After cleavage of the AP site by APE1, repair proceeds through re-synthesis and ligation steps. In mammalian cells, the XRCC1 protein, essential for the maintenance of genomic stability, is involved in both base excision and single-strand break repair. XRCC1 participates in the first step of BER by interacting with the human DNA glycosylases hOGG1 and NEIL1. To analyze the possibility of a general mechanism involving the interaction of XRCC1 with DNA glycosylases we used XRCC1 to pull-down DNA glycosylases activities from human cell extracts. XRCC1 co-purifies with DNA glycosylase activities capable of excising hypoxanthine and dihydrothymine, in addition to 8-oxoguanine, but not uracil. Biochemical analyses with the purified proteins confirmed the interactions between XRCC1 and MPG, hNTH1 or hNEIL2. Furthermore, XRCC1 stimulates the activities of these enzymes. In vivo localization studies show that after genotoxic treatments these DNA glycosylases can be found associated with XRCC1 foci. Our results support a BER model in which XRCC1 is recruited to the repair of alkylated or oxidized bases by the enzyme recognizing the lesion. XRCC1 would then coordinate the subsequent enzymatic steps and modulate the activities of all the proteins involved.
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U2 - 10.1016/j.dnarep.2005.04.014
DO - 10.1016/j.dnarep.2005.04.014
M3 - Article
C2 - 15927541
AN - SCOPUS:20644453965
SN - 1568-7864
VL - 4
SP - 826
EP - 835
JO - DNA Repair
JF - DNA Repair
IS - 7
ER -