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Genetics, Vol 123, 123-129, Copyright © 1989
INVESTIGATIONS |
Sequence Analysis of N-Ethyl-N-Nitrosourea-Induced vermilion Mutations in Drosophila melanogaster
A. Pastink, C. Vreeken, MJM. Nivard, L. L. Searles and E. W. Vogel
Department of Radiation Genetics and Chemical Mutagenesis, State University of Leiden, Sylvius Laboratories, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands, J. A. Cohen Institute, Interuniversity Research Institute for Radiopathology and Radiation Protection, The Netherlands
The mutational specificity of N-ethyl-N-nitrosourea (ENU) was determined in Drosophila melanogaster using the vermilion locus as a target gene. 25 mutants (16 F(1) and 9 F(2) mutants) were cloned and sequenced. Only base-pair changes were observed; three of the mutants represented double base substitutions. Transition mutations were the most prominent sequence change: 61% were GC->AT and 18% AT->GC substitutions. Both sequence changes can be explained by the miscoding properties of the modified guanine and thymine bases. A strong bias of neighboring bases on the occurrence of the GC->AT transitions or a strand preference of both types of transition mutations was not observed. The spectrum of ENU mutations in D. melanogaster includes a significant fraction (21%) of transversion mutations. Our data indicate that like in other prokaryotic and eukaryotic systems also in D. melanogaster the O(6)-ethylguanine adduct is the most prominent premutational lesion after ENU treatment. The strong contribution of the O(6)-ethylguanine adduct to the mutagenicity of ENU possibly explains the absence of distinct differences between the type of mutations observed in the F(1) and F(2) mutants. Although the latter arise later during development, the spectrum of mosaic mutations is also dominated by GC->AT transition mutations.
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