Simultaneous measurement of the frequencies of intrachromosomal recombination and chromosome gain using the yeast DEL assay.
The yeast DEL assay measures the frequency of intrachromosomal recombination between two partially-deleted his3 alleles on chromosome XV. The his3Delta alleles share approximately 400bp of overlapping homology, and are separated by an intervening LEU2 sequence. Homologous recombination between the his3Delta alleles results in deletion of the intervening LEU2 sequence (DEL), and reversion to histidine prototrophy. In this study we have attempted to further extend the use of the yeast DEL assay to measure the frequency of chromosome XV gain events. Reversion to His(+)Leu(+) in the haploid yeast DEL tester strain RSY6 occurs upon non-disjunction of chromosome XV sister chromatids, coupled with a subsequent DEL event. Here we have tested the ability of the yeast DEL assay to accurately predict the aneugenic potential of the diversely-acting, known or suspected aneugens actinomycin D, benomyl, chloral hydrate, ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), and methotrexate. Actinomycin D and benomyl strongly induced aneuploidy. EMS and methotrexate modestly induced aneuploidy, while chloral hydrate and MMS failed to illicit any significant induction. In addition, by FACS-analysis of DNA content it was shown that the majority of both spontaneous- and chemically-induced His(+)Leu(+) revertants were heterodiploid. Thus, our results indicate endoreduplication of almost entire chromosome sets as a major mechanism of aneuploidy induction in haploid Saccharomyces cerevisiae.
Howlett
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Department of Cancer Cell Biology, Harvard School of Public Health, 665 Huntington Avenue, Boston, MA 02115, USA.
Schiestl
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RH
D000782:Aneuploidy; D001542:Benomyl / Q000494:pharmacology; D002697:Chloral Hydrate / Q000494:pharmacology; D015825:Chromosomes, Fungal; D003609:Dactinomycin / Q000494:pharmacology; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D005434:Flow Cytometry; D008727:Methotrexate / Q000494:pharmacology; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009153:Mutagens / Q000494:pharmacology; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D009153:Mutagens; D003609:Dactinomycin; D002697:Chloral Hydrate; D005020:Ethyl Methanesulfonate; D008741:Methyl Methanesulfonate; D001542:Benomyl; D008727:Methotrexate