Several spontaneous Mn2+-resistant mutants were isolated from Saccharomyces cerevisiae strain W303-1b. All displayed an identical semidominant resistance phenotype. The gene responsible for this phenotype from one of these mutants, as well as that from the wild-type, was cloned and sequenced, which allowed the identification of a single nucleotide change in the former. Further sequencing work with the remaining mutants, as well as with others on the MNAR1 gene (Bianchi et al., 1981), indicated that all displayed the same mutation (mnr1 allele). In addition, MNR1 was shown to correspond to VCX1 and HUM1, which determine a vacuolar membrane protein. The nucleotide sequence has been deposited at EMBL, with Accession No. AJ001272.