Repair of a synthetic abasic site involves concerted reactions of DNA synthesis followed by excision and ligation.
Open Access
- 1 September 1991
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 11 (9) , 4441-4447
- https://doi.org/10.1128/mcb.11.9.4441
Abstract
A synthetic analog of an abasic site in DNA is efficiently repaired by a short-patch repair mechanism in soluble extracts of Xenopus laevis oocytes (Y. Matsumoto and D. F. Bogenhagen, Mol. Cell. Biol. 9:3750-3757, 1989). We present a detailed analysis of the repair mechanism, using extracts depleted of endogenous nucleotide pools. ATP was required for repair with a sharp optimal concentration of 5 mM. The initial rate of repair was increased by preincubation of the DNA in the extract in the presence of ATP. During this preincubation, the DNA was cleaved on the 5' side of the lesion by a class II apurinic-apyrimidinic endonuclease, but removal of the abasic sugar residue was not observed prior to addition of deoxynucleotides to the reaction. Immediately following DNA synthesis, excision and ligation proceeded in a coordinated manner to complete repair. DNA preincubated in the extract in the absence of deoxynucleotides remained associated with repair enzymes during gel filtration. These observations suggest that the enzymes involved in concerted repair of the abasic site form a complex on DNA.Keywords
This publication has 14 references indexed in Scilit:
- Heteroduplex repair in extracts of human HeLa cellsJournal of Biological Chemistry, 1991
- DNA mismatch repair in Xenopus egg extracts: repair efficiency and DNA repair synthesis for all single base-pair mismatches.Proceedings of the National Academy of Sciences, 1990
- Mismatch-specific thymine DNA glycosylase and DNA polymerase beta mediate the correction of G.T mispairs in nuclear extracts from human cells.Proceedings of the National Academy of Sciences, 1990
- Complementation of the xeroderma pigmentosum DNA repair synthesis defect withEscherichia coliUvrABC proteins in a cell-free systemNucleic Acids Research, 1990
- Localization of DNA repair synthesis by human cell extracts to a short region at the site of a lesionJournal of Biological Chemistry, 1989
- Oligodeoxynucleotides containing synthetic abasic sites. Model substrates for DNA polymerases and apurinic/apyrimidinic endonucleases.Journal of Biological Chemistry, 1987
- Apurinic sites as mutagenic intermediatesCell, 1985
- Chromatin assembly in Xenopus oocytes: In vitro studiesCell, 1984
- A simple, rapid, and sensitive DNA assay procedureAnalytical Biochemistry, 1980
- Rate of depurination of native deoxyribonucleic acidBiochemistry, 1972