title: Radiation affects binding of Fpg repair protein to an abasic site containing DNA.
abstract: During the base excision repair of certain DNA lesions, the formamidopyrimidine-DNA glycosylase (Fpg) binds specifically to the DNA region containing an abasic (AP) site. Is this step affected by exposure to ionizing radiation? To answer this question, we studied a complex between a DNA duplex containing an analogue of an abasic site (the 1,3-propanediol site, Pr) and a mutated Lactococcus lactis Fpg (P1G-LlFpg) lacking strand cleavage activity. Upon irradiation of the complex, the ratio of bound/free partners decreased. When the partners were irradiated separately, the irradiated DNA still bound the unirradiated protein, whereas irradiated Fpg no longer bound unirradiated DNA. Thus irradiation hinders Fpg-DNA binding because of the damage to the protein. Using our radiolytic attack simulation procedure RADACK (Begusova et al., J. Biomol. Struct. Dyn. 19, 141-157, 2001), we reveal the potential hot spots for damage in the irradiated protein. Most of them are essential for the interaction of Fpg with DNA, which explains the radiation-induced loss of binding ability of Fpg. The doses necessary to destroy the complex are higher than those inactivating Fpg irradiated separately. As confirmed by our calculations, this can be explained by the partial protection of the protein by the bound DNA.
authors.affiliation: Centre de Biophysique Moléculaire CNRS, F-45071 Orléans Cedex 2, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Gillard
authors.forename: Marie
authors.initials: M
authors.lastname: Begusova
authors.forename: Bertrand
authors.initials: B
authors.lastname: Castaing
authors.forename: Melanie
authors.initials: M
authors.lastname: Spotheim-Maurizot
mesh_terms: D004247:DNA / Q000378:metabolism / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair; D043382:DNA-Formamidopyrimidine Glycosylase / Q000737:chemistry / Q000378:metabolism*; D004307:Dose-Response Relationship, Radiation; D013294:Lactococcus lactis / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding
chemical_list: D004247:DNA; D043382:DNA-Formamidopyrimidine Glycosylase