Mitotic recombination and localized DNA double-strand breaks are induced after 8-methoxypsoralen and UVA irradiation in Saccharomyces cerevisiae.
Mitotic recombination within the ARG4 gene of Saccharomyces cerevisiae was analysed after treatment of cells with the recombinogenic agent 8-methoxypsoralen (8-MOP) plus UVA. The appearance of DNA double-strand breaks (DSBs) in the ARG4 region during post-treatment incubation was also tested. The results obtained after 8-MOP plus UVA treatment indicate that in mitotic cells: (1) recombination at the ARG4 locus is increased 30 - 500 fold per survivor depending on the strains and the doses employed, (2) the increase of recombination results essentially from gene conversion events which involve the RV site located in the 5' region of the ARG4 gene twice as often as the Bgl site at the 3' end, (3) depending on 8-MOP/UVA dose, ectopic gene conversion is associated with reciprocal translocation, (4) DSBs occur preferentially in the ARG 5' region during post-treatment incubation, as well as in other intergenic regions containing both promoters or/and terminators of transcription, and (5) changes in sequence content in the 5' region of ARG4, which influences positions and frequencies of DSBs formed during repair, are correlated with a modification of the local chromatin structure.
Institut Curie section de recherche, UMR 218 CNRS, LRC no. I du CEA, 26, rue d'Ulm, F-75248 Paris cedex 05, France. Michele.Dardalhon@curie.fr
M
M
Dardalhon
B
B
de Massy
A
A
Nicolas
D
D
Averbeck
D001123:Argininosuccinate Lyase; D015139:Blotting, Southern; D003434:Crossing Over, Genetic; D004249:DNA Damage; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004271:DNA, Fungal / Q000187:drug effects / Q000528:radiation effects; D005656:Fungal Proteins / Q000235:genetics*; D005785:Gene Conversion; D008730:Methoxsalen / Q000494:pharmacology*; D008938:Mitosis / Q000235:genetics*; D017319:Photosensitizing Agents / Q000494:pharmacology; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins; D014466:Ultraviolet Rays
D004271:DNA, Fungal; D005656:Fungal Proteins; D017319:Photosensitizing Agents; D029701:Saccharomyces cerevisiae Proteins; D004262:DNA Restriction Enzymes; C087900:Arg4 protein, S cerevisiae; D001123:Argininosuccinate Lyase; D008730:Methoxsalen