Reconstitution of proliferating cell nuclear antigen-dependent repair of apurinic/apyrimidinic sites with purified human proteins Journal Article
| Authors: | Matsumoto, Y.; Kim, K.; Hurwitz, J.; Gary, R.; Levin, D. S.; Tomkinson, A. E.; Park, M. S. |
| Article Title: | Reconstitution of proliferating cell nuclear antigen-dependent repair of apurinic/apyrimidinic sites with purified human proteins |
| Abstract: | An apurinic/apyrimidinic (AP) site is one of the most abundant lesions spontaneously generated in living cells and is also a reaction intermediate in base excision repair. In higher eukaryotes, there are two alternative pathways for base excision repair: a DNA polymerase β-dependent pathway and a proliferating cell nuclear antigen (PCNA)-dependent pathway. Here we have reconstituted PCNA-dependent repair of AP sites with six purified human proteins: AP endonuclease, replication factor C, PCNA, flap endonuclease 1 (FEN1), DNA polymerase δ, and DNA ligase I. The length of nucleotides replaced during the repair reaction (patch size) was predominantly two nucleotides, although longer patches of up to seven nucleotides could be detected. Neither replication protein A nor Ku70/80 enhanced the repair activity in this system. Disruption of the PCNA-binding site of either FEN1 or DNA ligase I significantly reduced efficiency of AP site repair but did not affect repair patch size. |
| Keywords: | dna polymerase; dna replication; dna synthesis; proteins; dna repair; protein protein interaction; eukaryota; base sequence; binding site; excision repair; dna primers; cycline; flap endonucleases; polydeoxyribonucleotide synthase; enzyme binding; replication protein a; dna ligases; exodeoxyribonucleases; proliferating cell nuclear antigen; exodeoxyribonuclease v; dna directed dna polymerase beta; enzyme reconstitution; humans; human; priority journal; article; dna (apurinic or apyrimidinic site) lyase; apurinic site; apyrimidinic site |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 274 |
| Issue: | 47 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1999-11-19 |
| Start Page: | 33703 |
| End Page: | 33708 |
| Language: | English |
| DOI: | 10.1074/jbc.274.47.33703 |
| PUBMED: | 10559261 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 16 August 2016 -- Source: Scopus |
