In Schizosaccharomyces pombe the frequency of mating-type (MT) switching is reduced by mutations in the swi genes. The ten hitherto known swi genes can be subdivided into three classes: Ia, Ib and II. Strains having swi5 (class Ib), swi9 (class II) and swi10 (class II) mutations do not only show reduced MT switching, but also exhibit an increased sensitivity to UV- and gamma-rays. For that reason, 19 previously described rad genes were tested for their effect on MT switching. We found that swi9, "rad10", "rad16" and "rad20" are allelic with each other indicating that the former allocation of these rad mutations to three different genes must have been erroneous. Among the remaining 16 rad genes examined, rad22 seems to be a new class II swi gene. The double mutants swi5 swi9 and swi5 swi10, but not swi9 swi10, are much more sensitive to radiation than the respective single mutants. Thus a cumulative increase in sensitivity occurs only if the mutants belong to different classes; previously the same correlation was found with regard to cumulative effects in MT switching.
Mutanase (Mutanase Novo) affects the high frequency production of protoplasts in the following strains of yeast: Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Trichosporon pullulans, and Schwanniomyces alluvius. Regeneration frequencies varied with the strain used and ranged between 10 and 18%. This enzyme preparation appears to be a very useful means of obtaining protoplasts from a wide variety of yeasts currently being used for experimental purposes.
A method is presented to screen chemicals for potential mutagenicity on the basis of their ability to cause more killing in cells of repair-deficient yeast than in wild type cells. Two species were chosen in the event that one might be more sensitive to certain chemicals. The strains used were RAD+ and rad6 derivatives of Saccharomyces cerevisiae and RAD+ and rad3 derivatives of Schizosaccharomyces pombe. This report describes the test system and results for 12 known, direct-acting mutagens (i.e., not requiring mammalian metabolic activation). These compounds showed more lethality in one or both of the repair-deficient strains, indicating that they induce damage to DNA which is subject to repair in wild type cells. Advantages of this system include the use of eukaryotic yeast cells which can be manipulated as easily as bacteria, and that exogenous enzymes (S9) can be added for metabolic activation. Growing yeast cells can activate certain promutagens, and preliminary experiments showed positive responses for diethylnitrosamine and 2-acetylaminofluorene without the addition of S9.
Following mutagenesis of yeast cells with nitrosoguanidine, primary mosaic colonies exhibiting prototrophic/auxotrophic phenotypes were obtained. Upon replating of these primary mosaics, numerous secondary mosaics were present in the progeny. This study shows that replicating instabilities occur at many different loci within the Schizosaccharomyces pombe genome. In addition, the ade-1 gene of Saccharomyces cerevisiae (causing red pigmentation) was used to show that the phenomenon also occurs in this yeast.
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