After ethylmethanesulfonate mutagenesis of the strain Saccharomyces cerevisiae D273-10B, out of 100,000 survivors, 1,000 were selected for their high production of petite mutants at 36 degrees C. Among these 1,000 mutators, 5 also showed an increased frequency of spontaneous point mutations measured at 25 degrees C. Further analysis revealed that in all mutators, except 2, petite accumulation proceeded at 25 degrees C as well as 36 degrees C. In these 2 mutants, the production of petite mutants was much higher at 36 degrees C than at 35 degrees C. In one of them, however, the mutator and the thermosensitive petite phenotypes were due to mutations in two unlinked nuclear genes. In the other mutants, both traits were the result of a mutation in a single nuclear gene. The mutators fell into three complementation groups (tpm1, tpm2, mup1). No complementation was observed between tpm1 mutants and the gam4 mutant previously described by Foury and Goffeau (1979). From the latter and the present works, only four complementation groups (gam1, gam2, gam4 or tpm1, mup1) have been identified and it is likely that the number of genes controlling specifically the spontaneous mutability of the mtDNA is low. The mutators exhibited a variety of responses to damaging agents such as UV light and ethidium bromide; especially in a representative mutant from the complementation group tpm1, the induction of rho- mutants was sensitive to UV light and resistant to ethidium bromide.(ABSTRACT TRUNCATED AT 250 WORDS)