To examine the interaction of mammalian base excision repair (BER) enzymes with DNA intermediates formed during BER, we used a novel photoaffinity labeling probe and mouse embryonic fibroblast cellular extracts. The probe was formed in situ, using an end-labeled oligonucleotide containing a synthetic abasic site; this site was incised by apurinic/apyrimidinic endonuclease creating a nick with 3′-hydroxyl and 5′-reduced sugar phosphate groups at the margins, and then a dNMP carrying a photoreactive adduct was added to the 3′-hydroxyl group. With near-UV light (312 nm) exposure of the extract/probe mixture, six proteins were strongly labeled. Four of these include poly(ADP-ribose) polymerase-1 (PARP-1) and the BER participants flap endonuclease-1, DNA polymerase β, and apurinic/apyrimidinic endonuclease. The amount of the probe cross-linked to PARP-1 was greater than that cross-linked to the other proteins. The specificity of PARP-1 labeling was examined using various competitor oligonucleotides and DNA probes with alternate structures. PARP-1 labeling was stronger with a DNA representing a BER intermediate than with a nick in double-stranded DNA. These results indicate that proteins interacting preferentially with a photoreactive BER intermediate can be selected from the crude cellular extract. To examine the interaction of mammalian base excision repair (BER) enzymes with DNA intermediates formed during BER, we used a novel photoaffinity labeling probe and mouse embryonic fibroblast cellular extracts. The probe was formed in situ, using an end-labeled oligonucleotide containing a synthetic abasic site; this site was incised by apurinic/apyrimidinic endonuclease creating a nick with 3′-hydroxyl and 5′-reduced sugar phosphate groups at the margins, and then a dNMP carrying a photoreactive adduct was added to the 3′-hydroxyl group. With near-UV light (312 nm) exposure of the extract/probe mixture, six proteins were strongly labeled. Four of these include poly(ADP-ribose) polymerase-1 (PARP-1) and the BER participants flap endonuclease-1, DNA polymerase β, and apurinic/apyrimidinic endonuclease. The amount of the probe cross-linked to PARP-1 was greater than that cross-linked to the other proteins. The specificity of PARP-1 labeling was examined using various competitor oligonucleotides and DNA probes with alternate structures. PARP-1 labeling was stronger with a DNA representing a BER intermediate than with a nick in double-stranded DNA. These results indicate that proteins interacting preferentially with a photoreactive BER intermediate can be selected from the crude cellular extract. base excision repair apurinic/apyrimidinic endonuclease DNA polymerase β flap endonuclease-1 poly(ADP-ribose) polymerase x-ray cross complementing factor 1 replication protein A 5′- deoxyribose phosphate 3-hydroxy-2-hydroxymethyltetrahydrofuran polyacrylamide gel electrophoresis exo-N-[β-(p-azidotetrafluorobenzamido)-ethyl]deoxycytidine 5′-triphosphate exo-N-[β-(p-azidotetrafluorobenzamido)-ethyl]deoxycytidine 5′-monophosphate mouse embryonic fibroblast(s) fetal bovine serum Dulbecco's modified Eagle's medium 3-aminobenzamide 4-amino-1,8-naphthalimide methyl methanesulfonate Base excision repair (BER)1 is one of the main strategies of the cell for defense against exogenous and endogenous genotoxic stress leading to single-base lesions in DNA (1Lindahl T. Annu. Rev. Biochem. 1982; 51: 61-87Crossref PubMed Scopus (693) Google Scholar, 2Lindahl T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar). Mouse embryonic fibroblasts (MEFs) rendered deficient in BER by gene deletion of a BER enzyme are hypersensitive to monofunctional DNA-methylating agents among other stressors, illustrating the importance of this repair system (3Ochs K. Sobol R.W. Wilson S.H. Kaina B. Cancer Res. 1999; 59: 1544-1551PubMed Google Scholar, 4Sobol R.W. Horton J.K. Kuhn R. Gu H. Singhal R.K. Prasad R. Rajewsky K. Wilson S.H. Nature. 1996; 379: 183-186Crossref PubMed Scopus (783) Google Scholar, 5Wilson S.H. Mutat. Res. 1998; 407: 203-215Crossref PubMed Scopus (264) Google Scholar). The current and generally accepted working model for mammalian BER involves two sub-pathways, each proceeding as a sequential process with several DNA and/or DNA-enzyme intermediates (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, A. P. T. J. 1996; PubMed Scopus Google Scholar, K. PubMed Scopus Google Scholar, Prasad R. Wilson S.H. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar). each of the BER sub-pathways, repair can be by base DNA to an abasic site and in by base and (1Lindahl T. Annu. Rev. Biochem. 1982; 51: 61-87Crossref PubMed Scopus (693) Google Scholar, T. B. PubMed Scopus Google Scholar, G. B. T. PubMed Scopus Google Scholar). the abasic site is an DNA the of the sugar is by the enzyme apurinic/apyrimidinic endonuclease and the of the sugar is by the deoxyribose phosphate of DNA polymerase β Mutat. Res. PubMed Scopus Google Scholar, K. Science. PubMed Scopus Google Scholar). The is by and then the nick is by a DNA the BER A. P. T. J. 1996; PubMed Scopus Google Scholar, R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar, R.K. Prasad R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). mammalian repair of and abasic to by this G. J. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, A. A. Wood R.D. T. 1999; Full Text Full Text PubMed Scopus Google Scholar). other for the sugar of the abasic site is the BER repair R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). involves and DNA to and as as in the S.H. Mutat. Res. 1998; 407: 203-215Crossref PubMed Scopus (264) Google Scholar, J. PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, PubMed Scopus Google Scholar). this is by the flap endonuclease-1 and the nick is by a DNA R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar, A. A. Wood R.D. T. 1999; Full Text Full Text PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar). The DNA of BER can be by other DNA (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, Sobol R.W. Wilson S.H. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, Prasad R. Wilson S.H. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, R. K. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, J.K. Prasad R. E. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, K. Biol. PubMed Scopus Google Scholar). The BER to for a of BER in cell that BER Prasad R. Wilson S.H. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google J.K. Prasad R. E. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google various in and BER be and and for x-ray factor 1 and DNA A. P. T. J. 1996; PubMed Scopus Google Scholar, Mutat. Res. PubMed Scopus Google and for and the Wilson B. A. PubMed Scopus Google PARP-1 F. G. J. G. PubMed Scopus Google and DNA R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar). the and for each of these are to be and the of the the and of BER are the to be in this we examined the interaction PARP-1 and BER is an enzyme that is to to DNA and to using as of PARP-1 is to to from site in Biochem. J. 1999; PubMed Scopus Google Scholar). PARP-1 in mammalian in several F. P. T. J. G. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, H. R. B. 1999; PubMed Scopus Google Scholar, J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, A. T. Science. 1998; PubMed Scopus Google Scholar). F. G. J. G. PubMed Scopus Google examined the of PARP-1 in and BER using of cell for the and/or PARP-1 other that from PARP-1 were in BER and strongly in BER to a of PARP-1 in and a for PARP-1 in BER F. G. J. G. PubMed Scopus Google Scholar). A for PARP-1 in BER of the DNA specificity of PARP-1 for in DNA T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, G. J. Biochem. Full Text PubMed Scopus Google and the of PARP-1 with the BER proteins and Mutat. Res. PubMed Scopus Google Scholar, F. G. J. G. PubMed Scopus Google Scholar, P. Res. 1996; PubMed Scopus Google Scholar). PARP-1 are to be hypersensitive to agents that lesions by BER F. E. G. J. J. G. 1999; PubMed Scopus Google Scholar, B. A. P. G. A. PubMed Scopus Google Scholar). was that PARP-1 can DNA during repair in Nature. 1982; PubMed Scopus Google and was that and DNA a BER that can the for DNA of the nick J. Biol. Chem. Full Text Full Text PubMed Google Scholar). is that PARP-1 a in BER, the of this we examined the of proteins in crude can be by a photoaffinity DNA probe representing an intermediate in that six proteins in the crude were strongly by this BER several BER The results several of BER enzymes and BER of the various enzymes in mammalian BER is a of this we used a novel photoaffinity labeling to BER enzymes in the crude of the two BER sub-pathways, a DNA is a in BER and one can a DNA with abasic site as used as representing an intermediate at the to in BER at the to DNA in the BER (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). we of proteins to a DNA as a novel for cellular enzymes in The of a DNA with a photoreactive at the of the nick with near-UV light of photoreactive BER intermediate was in by using a carrying a photoreactive at the of the The is to be a for DNA P. H. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar). of the photoreactive the of a in creating the photoreactive BER intermediate the cell The DNA probe in of BER in the cell DNA DNA a in the is a for that the main for BER protein interaction with this DNA is the BER intermediate than the two of the DNA a nick with a with near-UV the DNA probe in be cross-linked to the DNA polymerase to other proteins of interacting with a DNA this as an of DNA probe DNA polymerase other cellular proteins was used in these cellular proteins interacting preferentially with a photoreactive BER intermediate can be selected from the proteins in the crude cellular extract. in the of of DNA and DNA enzymes in cellular that protein labeling with the cellular was as six proteins were strongly labeled. Four of these to be of the BER and These for were as the using various with and proteins of of were to be was in this is an cellular protein and is to be a DNA T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google Scholar, G. J. Biochem. Full Text PubMed Scopus Google Scholar). is that PARP-1 was a of labeling by the photoreactive BER intermediate used as probe in these that PARP-1 was by the protein and that several other BER proteins as DNA and and were labeled. of the labeling of PARP-1 and the that PARP-1 an as in BER, we to examine for PARP-1 labeling in PARP-1 labeling in the used in these several of labeling using photoreactive labeling as a of the of the DNA labeling was for a nick carrying an abasic site sugar phosphate at the that PARP-1 of this BER intermediate was and labeling be by oligonucleotides representing BER intermediates by double-stranded with an system of and each cross-linked to a of DNA interaction PARP-1 and and is in with the results by F. G. J. G. PubMed Scopus Google Scholar). These that PARP-1 with the of in the of DNA F. G. J. G. PubMed Scopus Google Scholar). used from deficient in and/or PARP-1 and that proteins are in the BER of abasic the of PARP-1 and BER were F. G. J. G. PubMed Scopus Google Scholar). The deletion of PARP-1 a BER in the cell the of the of PARP-1 in BER is T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google results the for PARP-1 of BER intermediates that the enzyme with DNA containing a sugar phosphate at the specificity of PARP-1 a sugar at the of a nick in a that PARP-1 a BER intermediate with this intermediate is formed in BER of a dNMP by leading to BER to a of PARP-1 in BER The is with the of in the cell used in these that a in repair of and we that this the of in BER of and cross-linked to the DNA probe using to the to These interaction for these BER proteins in a at a intermediate of is that PARP-1 to this BER intermediate the of BER, as the in BER and the excision in PARP-1 in this be an in the of BER and in is in to these Base excision repair (BER)1 is one of the main strategies of the cell for defense against exogenous and endogenous genotoxic stress leading to single-base lesions in DNA (1Lindahl T. Annu. Rev. Biochem. 1982; 51: 61-87Crossref PubMed Scopus (693) Google Scholar, 2Lindahl T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar). Mouse embryonic fibroblasts (MEFs) rendered deficient in BER by gene deletion of a BER enzyme are hypersensitive to monofunctional DNA-methylating agents among other stressors, illustrating the importance of this repair system (3Ochs K. Sobol R.W. Wilson S.H. Kaina B. Cancer Res. 1999; 59: 1544-1551PubMed Google Scholar, 4Sobol R.W. Horton J.K. Kuhn R. Gu H. Singhal R.K. Prasad R. Rajewsky K. Wilson S.H. Nature. 1996; 379: 183-186Crossref PubMed Scopus (783) Google Scholar, 5Wilson S.H. Mutat. Res. 1998; 407: 203-215Crossref PubMed Scopus (264) Google Scholar). The current and generally accepted working model for mammalian BER involves two sub-pathways, each proceeding as a sequential process with several DNA and/or DNA-enzyme intermediates (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, A. P. T. J. 1996; PubMed Scopus Google Scholar, K. PubMed Scopus Google Scholar, Prasad R. Wilson S.H. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar). each of the BER sub-pathways, repair can be by base DNA to an abasic site and in by base and (1Lindahl T. Annu. Rev. Biochem. 1982; 51: 61-87Crossref PubMed Scopus (693) Google Scholar, T. B. PubMed Scopus Google Scholar, G. B. T. PubMed Scopus Google Scholar). the abasic site is an DNA the of the sugar is by the enzyme apurinic/apyrimidinic endonuclease and the of the sugar is by the deoxyribose phosphate of DNA polymerase β Mutat. Res. PubMed Scopus Google Scholar, K. Science. PubMed Scopus Google Scholar). The is by and then the nick is by a DNA the BER A. P. T. J. 1996; PubMed Scopus Google Scholar, R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar, R.K. Prasad R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). mammalian repair of and abasic to by this G. J. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, A. A. Wood R.D. T. 1999; Full Text Full Text PubMed Scopus Google Scholar). other for the sugar of the abasic site is the BER repair R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). involves and DNA to and as as in the S.H. Mutat. Res. 1998; 407: 203-215Crossref PubMed Scopus (264) Google Scholar, J. PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, PubMed Scopus Google Scholar). this is by the flap endonuclease-1 and the nick is by a DNA R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar, A. A. Wood R.D. T. 1999; Full Text Full Text PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar). The DNA of BER can be by other DNA (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, K. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, Sobol R.W. Wilson S.H. J. Biol. Chem. 1998; Full Text Full Text PubMed Scopus Google Scholar, Prasad R. Wilson S.H. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, R. K. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, J.K. Prasad R. E. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, K. Biol. PubMed Scopus Google Scholar). The BER to for a of BER in cell that BER Prasad R. Wilson S.H. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google J.K. Prasad R. E. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). The various in and BER be and and for x-ray factor 1 and DNA A. P. T. J. 1996; PubMed Scopus Google Scholar, Mutat. Res. PubMed Scopus Google and for and the Wilson B. A. PubMed Scopus Google PARP-1 F. G. J. G. PubMed Scopus Google and DNA R. Singhal R.K. Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar). the and for each of these are to be and the of the the and of BER are the to be in this we examined the interaction PARP-1 and BER PARP-1 is an enzyme that is to to DNA and to using as of PARP-1 is to to from site in Biochem. J. 1999; PubMed Scopus Google Scholar). PARP-1 in mammalian in several F. P. T. J. G. J. Biol. Chem. 1999; Full Text Full Text PubMed Scopus Google Scholar, H. R. B. 1999; PubMed Scopus Google Scholar, J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar, A. T. Science. 1998; PubMed Scopus Google Scholar). F. G. J. G. PubMed Scopus Google examined the of PARP-1 in and BER using of cell for the and/or PARP-1 other that from PARP-1 were in BER and strongly in BER to a of PARP-1 in and a for PARP-1 in BER F. G. J. G. PubMed Scopus Google Scholar). A for PARP-1 in BER of the DNA specificity of PARP-1 for in DNA T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, G. J. Biochem. Full Text PubMed Scopus Google and the of PARP-1 with the BER proteins and Mutat. Res. PubMed Scopus Google Scholar, F. G. J. G. PubMed Scopus Google Scholar, P. Res. 1996; PubMed Scopus Google Scholar). PARP-1 are to be hypersensitive to agents that lesions by BER F. E. G. J. J. G. 1999; PubMed Scopus Google Scholar, B. A. P. G. A. PubMed Scopus Google Scholar). was that PARP-1 can DNA during repair in Nature. 1982; PubMed Scopus Google and was that and DNA a BER that can the for DNA of the nick J. Biol. Chem. Full Text Full Text PubMed Google Scholar). is that PARP-1 a in BER, the of this we examined the of proteins in crude can be by a photoaffinity DNA probe representing an intermediate in that six proteins in the crude were strongly by this BER several BER The results several of BER enzymes and BER of the various enzymes in mammalian BER is a of this we used a novel photoaffinity labeling to BER enzymes in the crude of the two BER sub-pathways, a DNA is a in BER and one can a DNA with abasic site as used as representing an intermediate at the to in BER at the to DNA in the BER (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). we of proteins to a DNA as a novel for cellular enzymes in The of a DNA with a photoreactive at the of the nick with near-UV light of photoreactive BER intermediate was in by using a carrying a photoreactive at the of the The is to be a for DNA P. H. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar). of the photoreactive the of a in creating the photoreactive BER intermediate the cell The DNA probe in of BER in the cell DNA DNA a in the is a for that the main for BER protein interaction with this DNA is the BER intermediate than the two of the DNA a nick with a with near-UV the DNA probe in be cross-linked to the DNA polymerase to other proteins of interacting with a DNA this as an of DNA probe DNA polymerase other cellular proteins was used in these cellular proteins interacting preferentially with a photoreactive BER intermediate can be selected from the proteins in the crude cellular extract. in the of of DNA and DNA enzymes in cellular that protein labeling with the cellular was as six proteins were strongly labeled. Four of these to be of the BER and These for were as the using various with and proteins of of were to be was in this is an cellular protein and is to be a DNA T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google Scholar, G. J. Biochem. Full Text PubMed Scopus Google Scholar). is that PARP-1 was a of labeling by the photoreactive BER intermediate used as probe in these that PARP-1 was by the protein and that several other BER proteins as DNA and and were labeled. of the labeling of PARP-1 and the that PARP-1 an as in BER, we to examine for PARP-1 labeling in PARP-1 labeling in the used in these several of labeling using photoreactive labeling as a of the of the DNA labeling was for a nick carrying an abasic site sugar phosphate at the that PARP-1 of this BER intermediate was and labeling be by oligonucleotides representing BER intermediates by double-stranded with an system of and each cross-linked to a of DNA interaction PARP-1 and and is in with the results by F. G. J. G. PubMed Scopus Google Scholar). These that PARP-1 with the of in the of DNA F. G. J. G. PubMed Scopus Google Scholar). used from deficient in and/or PARP-1 and that proteins are in the BER of abasic the of PARP-1 and BER were F. G. J. G. PubMed Scopus Google Scholar). The deletion of PARP-1 a BER in the cell the of the of PARP-1 in BER is T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google results the for PARP-1 of BER intermediates that the enzyme with DNA containing a sugar phosphate at the specificity of PARP-1 a sugar at the of a nick in a that PARP-1 a BER intermediate with this intermediate is formed in BER of a dNMP by leading to BER to a of PARP-1 in BER The is with the of in the cell used in these that a in repair of and we that this the of in BER of and cross-linked to the DNA probe using to the to These interaction for these BER proteins in a at a intermediate of is that PARP-1 to this BER intermediate the of BER, as the in BER and the excision in PARP-1 in this be an in the of BER and in is in to these The of the various enzymes in mammalian BER is a of this we used a novel photoaffinity labeling to BER enzymes in the crude of the two BER sub-pathways, a DNA is a in BER and one can a DNA with abasic site as used as representing an intermediate at the to in BER at the to DNA in the BER (6Frosina G. Fortini P. Rossi O. Carrozzino F. Raspaglio G. Cox L.S. Lane D.P. Abbondandolo A. Dogliotti E. J. Biol. Chem. 1996; 271: 9573-9578Abstract Full Text Full Text PubMed Scopus Google Scholar, R. Wilson S.H. J. Biol. Chem. Full Text Full Text PubMed Scopus Google Scholar). we of proteins to a DNA as a novel for cellular enzymes in The of a DNA with a photoreactive at the of the nick with near-UV light of photoreactive BER intermediate was in by using a carrying a photoreactive at the of the The is to be a for DNA P. H. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Prasad R. Wood Wilson S.H. J. Biol. Chem. 1996; 271: Full Text Full Text PubMed Scopus Google Scholar). of the photoreactive the of a in creating the photoreactive BER intermediate the cell The DNA probe in of BER in the cell DNA DNA a in the is a for that the main for BER protein interaction with this DNA is the BER intermediate than the two of the DNA a nick with a group. with near-UV the DNA probe in be cross-linked to the DNA polymerase to other proteins of interacting with a DNA this as an of DNA probe DNA polymerase other cellular proteins was used in these cellular proteins interacting preferentially with a photoreactive BER intermediate can be selected from the proteins in the crude cellular extract. in the of of DNA and DNA enzymes in cellular that protein labeling with the cellular was as six proteins were strongly labeled. Four of these to be of the BER and These for were as the using various with and proteins of of were to be was in this PARP-1 is an cellular protein and is to be a DNA T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google Scholar, G. J. Biochem. Full Text PubMed Scopus Google Scholar). is that PARP-1 was a of labeling by the photoreactive BER intermediate used as probe in these that PARP-1 was by the protein and that several other BER proteins as DNA and and were labeled. of the labeling of PARP-1 and the that PARP-1 an as in BER, we to examine for PARP-1 labeling in PARP-1 labeling in the used in these several of labeling using photoreactive labeling as a of the of the DNA labeling was for a nick carrying an abasic site sugar phosphate at the that PARP-1 of this BER intermediate was and labeling be by oligonucleotides representing BER intermediates by double-stranded DNA. with an system of and each cross-linked to a of DNA interaction PARP-1 and and is in with the results by F. G. J. G. PubMed Scopus Google Scholar). These that PARP-1 with the of in the of DNA F. G. J. G. PubMed Scopus Google Scholar). used from deficient in and/or PARP-1 and that proteins are in the BER of abasic the of PARP-1 and BER were F. G. J. G. PubMed Scopus Google Scholar). The deletion of PARP-1 a BER in the cell the of the of PARP-1 in BER is T. Wood R.D. Science. 1999; 286: 1897-1905Crossref PubMed Scopus (1272) Google Scholar, Biochem. J. 1999; PubMed Scopus Google Scholar). results the for PARP-1 of BER intermediates that the enzyme with DNA containing a sugar phosphate at the specificity of PARP-1 a sugar at the of a nick in a that PARP-1 a BER intermediate with this intermediate is formed in BER of a dNMP by leading to BER to a of PARP-1 in BER The is with the of in the cell used in these that a in repair of and we that this the of in BER of and cross-linked to the DNA probe using to the to These interaction for these BER proteins in a at a intermediate of is that PARP-1 to this BER intermediate the of BER, as the in BER and the excision in PARP-1 in this be an in the of BER and in is in to these are to for the photoreactive and for for and of this and for of this
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