Characterization of postreplication repair in mutagen sensitive strains of Drosophila melanogaster

Abstract

Mutants of Drosophila melanogaster, with suspected repair deficiencies, were analyzed for their capacity to repair damage induced by Xrays and UV radiation. Analysis was performed on cell cultures derived from embryos of homozygous mutant stocks. Postreplication repair following UV radiation has been analyzed in mutant stocks derived from a total of 10 complementation groups. Cultures were irradiated, pulse labeled, and incubated in the dark prior to analysis by alkaline sucrose gradient centrifugation. Kinetics of the molecular weight increase in newly synthesized DNA were assayed after cells had been incubated in the presence or absence of caffeine. Two separate pathways of postreplication repair have been tentatively identified by mutants derived from 4 complementation groups. The proposed caffeine sensitive pathway (CAS) is defined by mutants which also disrupt meiosis. The second pathway (CIS) is caffeine insensitive and is not yet associated with meiotic functions. All mutants deficient in postreplication repair are also sensitive to nitrogen mustard. The mutants investigated display a normal capacity to repair single strand breaks induced in DNA by Xrays, although 2 may possess a reduced capacity to repair damage caused by localized incorporation of high specific activity thymidine 3H. The data have been employed to construct a model for repair of UV induced damage in Drosophila DNA. Implications of the model for DNA repair in mammals are discussed.