Identifying sites of simultaneous DNA replication in eukaryotes by N-methyl-N'-nitro-N-nitrosoguanidine multiple mutagenesis.
N-methyl-N'-nitro-N-nitrosoguanidine (NG) induces certain classes of multiple mutations in yeast at high frequency. By selecting for mutation at one locus (his4 or leu1) one frequently obtains double mutants where another mutation to temperature sensitivity has also been induced. This multiple mutagenesis exhibits a considerable specificity: for mutation at one particular locus there is a high chance that another mutation will be found in the same cell at one of a restricted number of other loci. For any given locus (e.g. his4) there is a spectrum of sites at which temperature-sensitivity mutations are co-induced. This spectrum differs for different loci, such that the spectrum of sites co-mutating with leu1 differs completely from that for sites co-mutating with his4. This NG'induced co-mutation is interpreted in terms of NG acting to enhance mutagenesis at sites of simultaneous DNA replication within the cell. The results so obtained indicate a very strict control over the order and timing of gene replication in Saccharomyces cerevisiae, and it is suggested that it is now possible to use NG double mutagenesis to try and locate origins of replication in yeast.
D004261:DNA Replication; D006639:Histidine / Q000378:metabolism; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology*; D009153:Mutagens; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012641:Selection, Genetic
D009153:Mutagens; D008769:Methylnitronitrosoguanidine; D006639:Histidine