Mutations in the dim-1 gene of Neurospora crassa reduce the level of DNA methylation.
Mutants that show reduced DNA methylation were identified in a mutant screen based on the assumptions that (i) the nucleoside analog 5-azacytidine (5-azaC) promotes the formation of potentially lethal DNA-methyltransferase adducts; (ii) reduction in DNA methyltransferase will decrease the sensitivity of cells to 5-azaC; and (iii) this potential selective advantage will be enhanced in mutants that are deficient in the repair of 5-azaC-induced DNA damage. Of fifteen potential repair mutants screened for sensitivity to 5-azaC, five (mus-9, mus-10, mus-11, mus-18, and uvs-3) showed moderately increased sensitivity and two (mus-20, mei-3) showed highly increased sensitivity. A mus-20 mutation was used to isolate three non-complementing methylation mutants. The mutations, named dim-1 (defective in methylation), reduced female fertility, reduced methylation by 40-50%, and altered patterns of methylation. In wild-type strains hypomethylation per se fails to alter methylation specificity. We demonstrate a growth-phase-dependent change in methylation patterns, detectable only in hypomethylated DNA from dim+ cultures. This may represent a growth-phase-dependent change in the relative amounts of distinct species of methyltransferase, one of which may be encoded by the dim-1 gene.
Foss
H M
HM
Institute of Molecular Biology, University of Oregon, Eugene 97403-1229, USA.
Roberts
C J
CJ
Selker
E U
EU
D001374:Azacitidine / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects; D015254:DNA Modification Methylases / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal / Q000235:genetics*; D005816:Genetic Complementation Test; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008957:Models, Genetic; D009154:Mutation; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity
D008741:Methyl Methanesulfonate; D015254:DNA Modification Methylases; D001374:Azacitidine