title: Efficiency of incision of an AP site within clustered DNA damage by the major human AP endonuclease.
abstract: A major DNA lesion induced by ionizing radiation and formed on removal of oxidized base lesions by various glycosylases is an apurinic/apyrimidinic site (AP site). The presence of an AP site within clustered DNA damage, induced following exposure to ionizing radiation or radiomimetic anticancer agents, may present a challenge to the repair machinery of the cell, if the major human AP endonuclease, HAP1, does not efficiently incise the AP site. In this study, specific oligonucleotide constructs containing an AP site located at several positions opposite to another damage [5,6-dihydrothymine (DHT), 8-oxoG, AP site, or various types of single strand breaks] on the complementary strand were used to determine the relative efficiency of the purified HAP1 protein in incising an AP site(s) from clustered DNA damage. A base damage (DHT and 8-oxoG) on the opposite strand has little or no influence on the rate of incision of an AP site by HAP1. In contrast, the presence of either a second AP site or various types of single strand breaks, when located one or three bases 3' to the base opposite to the AP site, has a strong inhibitory effect on the efficiency of incision of an AP site. The efficiency of binding of HAP1 to an AP site is reduced by approximately 1 order of magnitude if a single strand break (SSB) is located one or three bases 3' to the site opposite to the AP site on the complementary strand. If the AP site and either a SSB or a second AP site are located at any of the other positions relative to each other, a double strand break may result.
authors.affiliation: Medical Research Council Radiation and Genome Stability Unit, Harwell, Didcot, Oxon OX11 0RD, United Kingdom.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: David-Cordonnier
authors.forename: Siobhan M T
authors.initials: SM
authors.lastname: Cunniffe
authors.forename: Ian D
authors.initials: ID
authors.lastname: Hickson
authors.forename: Peter
authors.initials: P
authors.lastname: O'Neill
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D019757:Carbon-Oxygen Lyases / Q000737:chemistry* / Q000235:genetics*; D020134:Catalytic Domain; D004249:DNA Damage; D018076:DNA, Complementary / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D043223:Deoxyribonuclease IV (Phage T4-Induced); D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D006151:Guanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D012689:Sequence Homology, Nucleic Acid; D013936:Thymidine / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D000276:Adjuvants, Immunologic; D018076:DNA, Complementary; D029968:Escherichia coli Proteins; D009841:Oligonucleotides; D006151:Guanosine; C046215:8-hydroxyguanosine; C029949:5,6-dihydrothymidine; D043223:Deoxyribonuclease IV (Phage T4-Induced); C468972:endonuclease IV, E coli; D019757:Carbon-Oxygen Lyases; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D010100:Oxygen; D013936:Thymidine