A requirement for PARP-1 for the assembly or stability of XRCC1 nuclear foci at sites of oxidative DNA damage.

The molecular role of poly (ADP-ribose) polymerase-1 in DNA repair is unclear. Here, we show that the single-strand break repair protein XRCC1 is rapidly assembled into discrete nuclear foci after oxidative DNA damage at sites of poly (ADP-ribose) synthesis. Poly (ADP-ribose) synthesis peaks during a 10 min treatment with H2O2 and the appearance of XRCC1 foci peaks shortly afterwards. Both sites of poly (ADP-ribose) and XRCC1 foci decrease to background levels during subsequent incubation in drug-free medium, consistent with the rapidity of the single-strand break repair process. The formation of XRCC1 foci at sites of poly (ADP-ribose) was greatly reduced by mutation of the XRCC1 BRCT I domain that physically interacts with PARP-1. Moreover, we failed to detect XRCC1 foci in Adprt1-/- MEFs after treatment with H2O2. These data demonstrate that PARP-1 is required for the assembly or stability of XRCC1 nuclear foci after oxidative DNA damage and suggest that the formation of these foci is mediated via interaction with poly (ADP-ribose). These results support a model in which the rapid activation of PARP-1 at sites of DNA strand breakage facilitates DNA repair by recruiting the molecular scaffold protein, XRCC1.

[1]  M. West,et al.  XRCC1 keeps DNA from getting stranded. , 2000, Mutation research.

[2]  H. Vrieling,et al.  Molecular analysis of ethyl methanesulfonate-induced mutations at the hprt gene in the ethyl methanesulfonate-sensitive Chinese hamster cell line EM-C11 and its parental line CHO9. , 1994, Cancer research.

[3]  K. Caldecott,et al.  Central Role for the XRCC1 BRCT I Domain in Mammalian DNA Single-Strand Break Repair , 2002, Molecular and Cellular Biology.

[4]  J. Tainer,et al.  DNA-bound structures and mutants reveal abasic DNA binding by APE1 DNA repair and coordination , 2000, Nature.

[5]  D. Barnes,et al.  Reconstitution of DNA base excision‐repair with purified human proteins: interaction between DNA polymerase beta and the XRCC1 protein. , 1996, The EMBO journal.

[6]  V. Schreiber,et al.  Poly(ADP-ribose) Polymerase-1 (PARP-1) Is Required in Murine Cell Lines for Base Excision Repair of Oxidative DNA Damage in the Absence of DNA Polymerase β* , 2003, The Journal of Biological Chemistry.

[7]  S. Nocentini Rejoining kinetics of DNA single- and double-strand breaks in normal and DNA ligase-deficient cells after exposure to ultraviolet C and gamma radiation: an evaluation of ligating activities involved in different DNA repair processes. , 1999, Radiation research.

[8]  J. Pleschke,et al.  Poly(ADP-ribose) Binds to Specific Domains in DNA Damage Checkpoint Proteins* , 2000, The Journal of Biological Chemistry.

[9]  K. Caldecott,et al.  XRCC1 polypeptide interacts with DNA polymerase beta and possibly poly (ADP-ribose) polymerase, and DNA ligase III is a novel molecular 'nick-sensor' in vitro. , 1996, Nucleic acids research.

[10]  T. Lindahl,et al.  Down-regulation of DNA repair synthesis at DNA single-strand interruptions in poly(ADP-ribose) polymerase-1 deficient murine cell extracts. , 2002, DNA repair.

[11]  K. Caldecott Mammalian DNA single‐strand break repair: an X‐ra(y)ted affair , 2001, BioEssays : news and reviews in molecular, cellular and developmental biology.

[12]  Samuel H. Wilson,et al.  Cells deficient in DNA polymerase beta are hypersensitive to alkylating agent-induced apoptosis and chromosomal breakage. , 1999, Cancer research.

[13]  M. Masson,et al.  XRCC1 Is Specifically Associated with Poly(ADP-Ribose) Polymerase and Negatively Regulates Its Activity following DNA Damage , 1998, Molecular and Cellular Biology.

[14]  M. Weinfeld,et al.  XRCC1 Stimulates Human Polynucleotide Kinase Activity at Damaged DNA Termini and Accelerates DNA Single-Strand Break Repair , 2001, Cell.

[15]  L. Thompson,et al.  A Chinese hamster ovary cell mutant (EM-C11) with sensitivity to simple alkylating agents and a very high level of sister chromatid exchanges. , 1992, Mutagenesis.

[16]  Y. Kubota,et al.  Independent roles of XRCC1's two BRCT motifs in recovery from methylation damage. , 2003, DNA repair.

[17]  G. de Murcia,et al.  Poly(ADP-ribose) polymerase-1: what have we learned from the deficient mouse model? , 2000, Mutation research.

[18]  P. Dollé,et al.  Poly(ADP-ribose) Polymerase-2 (PARP-2) Is Required for Efficient Base Excision DNA Repair in Association with PARP-1 and XRCC1* , 2002, The Journal of Biological Chemistry.

[19]  J. Tainer,et al.  DNA-bound structures and mutants reveal abasic DNA binding by APE1 and DNA repair coordination [corrected]. , 2000, Nature.

[20]  Yong-jie Xu,et al.  Excision of C-4′-oxidized Deoxyribose Lesions from Double-stranded DNA by Human Apurinic/Apyrimidinic Endonuclease (Ape1 Protein) and DNA Polymerase β* , 1998, The Journal of Biological Chemistry.

[21]  D. Ly,et al.  Misregulation of gene expression in primary fibroblasts lacking poly(ADP-ribose) polymerase. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  V. Schreiber,et al.  Base excision repair is impaired in mammalian cells lacking Poly(ADP-ribose) polymerase-1. , 2000, Biochemistry.

[23]  J. Lamerdin,et al.  Construction of a functional cDNA clone of the hamsterERCC2 DNA repair and transcription gene , 1996, Somatic cell and molecular genetics.

[24]  B. Demple,et al.  Interaction of human apurinic endonuclease and DNA polymerase beta in the base excision repair pathway. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[25]  T. Ochiya,et al.  Poly(ADP-ribose) polymerase gene disruption conferred mice resistant to streptozotocin-induced diabetes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[26]  A. Tomkinson,et al.  An alternative splicing event which occurs in mouse pachytene spermatocytes generates a form of DNA ligase III with distinct biochemical properties that may function in meiotic recombination , 1997, Molecular and cellular biology.

[27]  Samuel H. Wilson,et al.  DNA Polymerase (cid:1) -mediated Long Patch Base Excision Repair POLY(ADP-RIBOSE) POLYMERASE-1 STIMULATES STRAND DISPLACEMENT DNA SYNTHESIS* , 2022 .

[28]  M. Flannery,et al.  Requirement for the Xrcc1 DNA base excision repair gene during early mouse development. , 1999, Developmental biology.

[29]  I. Hickson,et al.  XRCC1 coordinates the initial and late stages of DNA abasic site repair through protein–protein interactions , 2001, The EMBO journal.

[30]  K. Caldecott DNA Single-Strand Break Repair and Spinocerebellar Ataxia , 2003, Cell.

[31]  Samuel H. Wilson,et al.  Passing the baton in base excision repair , 2000, Nature Structural Biology.

[32]  A. Carrano,et al.  A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister-chromatid exchange. , 1982, Mutation research.

[33]  J D Tucker,et al.  Characterization of the XRCC1-DNA ligase III complex in vitro and its absence from mutant hamster cells. , 1995, Nucleic acids research.

[34]  Z. Herceg,et al.  Functions of poly(ADP-ribose) polymerase (PARP) in DNA repair, genomic integrity and cell death. , 2001, Mutation research.

[35]  T. Lindahl,et al.  XRCC1 protein interacts with one of two distinct forms of DNA ligase III. , 1997, Biochemistry.

[36]  A. Natarajan,et al.  A high yield of translocations parallels the high yield of sister chromatid exchanges in the CHO mutant EM9. , 1998, Mutation research.

[37]  A. Kuzminov Single-strand interruptions in replicating chromosomes cause double-strand breaks , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[38]  L. Thompson,et al.  An interaction between the mammalian DNA repair protein XRCC1 and DNA ligase III , 1994, Molecular and cellular biology.

[39]  Penny A. Johnson,et al.  A Cell Cycle-Specific Requirement for the XRCC1 BRCT II Domain during Mammalian DNA Strand Break Repair , 2000, Molecular and Cellular Biology.

[40]  A. Bürkle Physiology and pathophysiology of poly(ADP‐ribosyl)ation * , 2001, BioEssays : news and reviews in molecular, cellular and developmental biology.

[41]  T. Sugimura,et al.  The response of Parp knockout mice against DNA damaging agents. , 2000, Mutation research.

[42]  A. Spradling,et al.  Chromatin Loosening by Poly(ADP)-Ribose Polymerase (PARP) at Drosophila Puff Loci , 2003, Science.

[43]  K. Caldecott,et al.  Mutation of a BRCT domain selectively disrupts DNA single-strand break repair in noncycling Chinese hamster ovary cells. , 2000, Proceedings of the National Academy of Sciences of the United States of America.