Processing of UV Damage in Vitro by FEN-1 Proteins as Part of an Alternative DNA Excision Repair Pathway
- 24 March 1999
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 38 (15) , 4809-4817
- https://doi.org/10.1021/bi990105i
Abstract
Ultraviolet (UV) irradiation induces predominantly cyclobutane and (6-4) pyrimidine dimer photoproducts in DNA. Several mechanisms for repairing these mutagenic UV-induced DNA lesions have been identified. Nucleotide excision repair is a major pathway, but mechanisms involving photolyases and DNA glycosylases have also been characterized. Recently, a novel UV damage endonuclease (UVDE) was identified that initiates an excision repair pathway different from previously established repair mechanisms. Homologues of UVDE have been found in eukaryotes as well as in bacteria. In this report, we have used oligonucleotide substrates containing site-specific cyclobutane pyrimidine dimers and (6-4) photoproducts for the characterization of this UV damage repair pathway. After introduction of single-strand breaks at the 5‘ sides of the photolesions by UVDE, these intermediates became substrates for cleavage by flap endonucleases (FEN-1 proteins). FEN-1 homologues from humans, Saccharomyces cerevisiae, and Schizosaccharomyces pombe all cleaved the UVDE-nicked substrates at similar positions 3‘ to the photolesions. T4 endonuclease V-incised DNA was processed in the same way. Both nicked and flapped DNA substrates with photolesions (the latter may be intermediates in DNA polymerase-catalyzed strand displacement synthesis) were cleaved by FEN-1. The data suggest that the two enzymatic activities, UVDE and FEN-1, are part of an alternative excision repair pathway for repair of UV photoproducts.Keywords
This publication has 23 references indexed in Scilit:
- Synthesis and characterisation of oligodeoxynucleotides containing thio analogues of (6-4) pyrimidine—pyrimidinone photo-dimersJournal of Molecular Biology, 1998
- Human RAD2 Homolog 1 5′- to 3′-Exo/Endonuclease Can Efficiently Excise a Displaced DNA Fragment Containing a 5′-Terminal Abasic Lesion by Endonuclease ActivityPublished by Elsevier ,1996
- Mechanism of Tracking and Cleavage of Adduct-damaged DNA Substrates by the Mammalian 5′- to 3′-Exonuclease/Endonuclease RAD2 Homologue 1 or Flap Endonuclease 1Journal of Biological Chemistry, 1996
- DNA EXCISION REPAIRAnnual Review of Biochemistry, 1996
- Two Pathways for Base Excision Repair in Mammalian CellsJournal of Biological Chemistry, 1996
- Synthesis of a Phosphoramidite Coupling Unit of the Pyrimidine (6−4) Pyrimidone Photoproduct and Its Incorporation into OligodeoxynucleotidesJournal of the American Chemical Society, 1996
- Calf 5′ to 3′ Exo/Endonuclease Must Slide from a 5′ End of the Substrate to Perform Structure-specific CleavagePublished by Elsevier ,1995
- UV Light as a Footprinting Agent: Modulation of UV-induced DNA Damage by Transcription Factors Bound at the Promoters of Three Human GenesJournal of Molecular Biology, 1995
- Conditional Lethality of Null Mutations in RTH1 That Encodes the Yeast Counterpart of a Mammalian 5′- to 3′-Exonuclease Required for Lagging Strand DNA Synthesis in Reconstituted SystemsJournal of Biological Chemistry, 1995
- Mechanism of action of the Escherichia coli UvrABC nuclease: Clues to the damage recognition problemBioEssays, 1993