XRCC1 coordinates disparate responses and multiprotein repair complexes depending on the nature and context of the DNA damage

TY - JOUR

T1 - XRCC1 coordinates disparate responses and multiprotein repair complexes depending on the nature and context of the DNA damage

AU - Hanssen-Bauer, Audun

AU - Solvang-Garten, Karin

AU - Sundheim, Ottar

AU - Peña-Diaz, Javier

AU - Andersen, Sonja

AU - Slupphaug, Geir

AU - Krokan, Hans E

AU - Wilson, David M

AU - Akbari, Mansour

AU - Otterlei, Marit

N1 - Copyright © 2011 Wiley-Liss, Inc.

PY - 2011/10

Y1 - 2011/10

N2 - XRCC1 is a scaffold protein capable of interacting with several DNA repair proteins. Here we provide evidence for the presence of XRCC1 in different complexes of sizes from 200 to 1500 kDa, and we show that immunoprecipitates using XRCC1 as bait are capable of complete repair of AP sites via both short patch (SP) and long patch (LP) base excision repair (BER). We show that POLβ and PNK colocalize with XRCC1 in replication foci and that POLβ and PNK, but not PCNA, colocalize with constitutively present XRCC1-foci as well as damage-induced foci when low doses of a DNA-damaging agent are applied. We demonstrate that the laser dose used for introducing DNA damage determines the repertoire of DNA repair proteins recruited. Furthermore, we demonstrate that recruitment of POLβ and PNK to regions irradiated with low laser dose requires XRCC1 and that inhibition of PARylation by PARP-inhibitors only slightly reduces the recruitment of XRCC1, PNK, or POLβ to sites of DNA damage. Recruitment of PCNA and FEN-1 requires higher doses of irradiation and is enhanced by XRCC1, as well as by accumulation of PARP-1 at the site of DNA damage. These data improve our understanding of recruitment of BER proteins to sites of DNA damage and provide evidence for a role of XRCC1 in the organization of BER into multiprotein complexes of different sizes.

AB - XRCC1 is a scaffold protein capable of interacting with several DNA repair proteins. Here we provide evidence for the presence of XRCC1 in different complexes of sizes from 200 to 1500 kDa, and we show that immunoprecipitates using XRCC1 as bait are capable of complete repair of AP sites via both short patch (SP) and long patch (LP) base excision repair (BER). We show that POLβ and PNK colocalize with XRCC1 in replication foci and that POLβ and PNK, but not PCNA, colocalize with constitutively present XRCC1-foci as well as damage-induced foci when low doses of a DNA-damaging agent are applied. We demonstrate that the laser dose used for introducing DNA damage determines the repertoire of DNA repair proteins recruited. Furthermore, we demonstrate that recruitment of POLβ and PNK to regions irradiated with low laser dose requires XRCC1 and that inhibition of PARylation by PARP-inhibitors only slightly reduces the recruitment of XRCC1, PNK, or POLβ to sites of DNA damage. Recruitment of PCNA and FEN-1 requires higher doses of irradiation and is enhanced by XRCC1, as well as by accumulation of PARP-1 at the site of DNA damage. These data improve our understanding of recruitment of BER proteins to sites of DNA damage and provide evidence for a role of XRCC1 in the organization of BER into multiprotein complexes of different sizes.

KW - Animals

KW - Blotting, Western

KW - CHO Cells

KW - Cell Culture Techniques

KW - Cricetinae

KW - Cricetulus

KW - DNA Breaks, Single-Stranded

KW - DNA Polymerase beta

KW - DNA Repair

KW - DNA-Binding Proteins

KW - Dose-Response Relationship, Radiation

KW - HeLa Cells

KW - Humans

KW - Immunoprecipitation

KW - Lasers

KW - Microscopy, Confocal

KW - Models, Biological

KW - Poly(ADP-ribose) Polymerases

KW - Polynucleotide 5'-Hydroxyl-Kinase

KW - Proliferating Cell Nuclear Antigen

KW - Recombinant Fusion Proteins

KW - Transfection

U2 - 10.1002/em.20663

DO - 10.1002/em.20663

M3 - Journal article

C2 - 21786338

SN - 0898-3003

VL - 52

SP - 623

EP - 635

JO - Environmental and Molecular Mutagenesis

JF - Environmental and Molecular Mutagenesis

IS - 8

ER -