In vitro reconstitution of the Schizosaccharomyces pombe alternative excision repair pathway Journal Article
| Authors: | Alleva, J. L.; Zuo, S.; Hurwitz, J.; Doetsch, P. W. |
| Article Title: | In vitro reconstitution of the Schizosaccharomyces pombe alternative excision repair pathway |
| Abstract: | Schizosaccharomyces pombe alternative excision repair has been shown genetically and biochemically to be involved in the repair of a wide variety of DNA lesions. AER is initiated by a damage-specific endonuclease (Uve1p) that recognizes UV-induced photoproducts, base mispairs, abasic sites, and platinum G-G diadducts and cleaves the DNA phosphodiester backbone 5' to a lesion. Several models exist that employ various mechanisms for damage removal based on the activities of Rad2p, a nuclease thought to be responsible for damage excision in AER. This study represents the first report of the biochemical reconstitution of the AER pathway. A base mispair- containing substrate is repaired in a reaction requiring S. pombe Uve1p, Rad2p, DNA polymerase δ, replication factor C, proliferating cell nuclear antigen, and T4 DNA ligase. Surprisingly, damage is removed exclusively by the 5' to 3' exonuclease activity of Rad2p and not its 'flap endonuclease' activity and is absolutely dependent upon the presence of the 5'-phosphoryl moiety at the Uve1p cleavage site. |
| Keywords: | dna-binding proteins; nonhuman; dna damage; dna repair; enzyme activation; substrate specificity; nuclease; excision repair; saccharomyces cerevisiae proteins; replication factor c; cycline; flap endonucleases; endonuclease; polydeoxyribonucleotide synthase; hydrolysis; schizosaccharomyces; schizosaccharomyces pombe proteins; schizosaccharomyces pombe; endodeoxyribonucleases; exodeoxyribonucleases; exodeoxyribonuclease v; base mispairing; fungal proteins; dna directed dna polymerase delta; priority journal; article |
| Journal Title: | Biochemistry |
| Volume: | 39 |
| Issue: | 10 |
| ISSN: | 0006-2960 |
| Publisher: | American Chemical Society |
| Date Published: | 2000-03-14 |
| Start Page: | 2659 |
| End Page: | 2666 |
| Language: | English |
| DOI: | 10.1021/bi992751n |
| PUBMED: | 10704216 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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