Exp Mol Med.
2003 Apr;35(2):106-112.
Base excision repair synthesis of DNA containing 8-oxoguanine in Escherichia coli
- Affiliations
-
- 1Division of Pharmacology, Sungkyunkwan University School of Medicine, Suwon 440-746, Korea. yslee@skku.ac.kr
- 2Department of Molecular and Cellular Biology Sungkyunkwan University School of Medicine, Suwon 440-746, Korea.
- 3Department of Pharmacology Seoul National University College of Medicine Jongno-gu, Seoul 110-799, Korea.
Abstract
- 8-oxo-7,8-dihydroguanine (8-oxo-G) in DNA is a mutagenic adduct formed by reactive oxygen species. In Escherichia coli, 2,6-dihydroxy-5N-formamidopyrimidine (Fapy)-DNA glycosylase (Fpg) removes this mutagenic adduct from DNA. In this report, we demonstrate base excision repair (BER) synthesis of DNA containing 8-oxo-G with Fpg in vitro. Fpg cut the oligonucleotide at the site of 8-oxo-G, producing one nucleotide gap with 3' and 5' phosphate termini. Next, 3' phosphatase(s) in the supernatant obtained by precipitating a crude extract of E. coli with 40% ammonium sulfate, removed the 3' phosphate group at the gap, thus exposing the 3' hydroxyl group to prime DNA synthesis. DNA polymerase and DNA ligase then completed the repair. These results indicate the biological significance of the glycosylase and apurinic/ apyrimidinic (AP) lyase activities of Fpg, in concert with 3' phosphatase(s) to create an appropriately gapped substrate for efficient BER synthesis of DNA containing 8-oxo-G.
Keyword
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Identification of Escherichia coli 8-oxoguanine endonuclease
- Escherichia coli O157: H7 Infection
- The potential roles of p53 tumor suppressor in nucleotide excision repair (NER) and base excision repair (BER)
- The Effect of Genetic Variation in The Dna Base Repair Genes on the Risk of Head and Neck Cancer
- Cloning and Sequencing of the phoA Gene which is Regulated by the phoP-phoQ operon in Pathogenic Enteric Bacteria
