Loss of heterozygosity, by mitotic gene conversion and crossing over, causes strain-specific adenine mutants in constitutive diploid Candida albicans

Abstract

Molecular evidence is provided in this paper to elucidate a long-standing intriguing phenomenon in fungal genetics: that many natural isolates of the constitutive diploid organism Candida albicans yield strain-specific, recessive mutants at a reproducible frequency that is as high as a few percent of the surviving cells after exposure to UV irradiation or other mutagens. Southern hybridization analysis and DNA sequence data indicated that C. albicans CA12, a clinical isolate, is heterozygous for the ADE2 gene, carrying one functional and one null allele. Sequence analysis of the null allele revealed the presence of a 1.3 kb deletion, which locates between two AATC repeats and spans the promoter and coding regions of the gene. The adenine auxotrophic mutants, which were readily isolated after UV irradiation of C. albicans CA12, were proved to be the segregants of mitotic recombination as they remained as diploid, not hemizygous or haploid, cells and were homozygous for ade2. Analysis of reciprocal products of the mitotic recombination detected that the process of loss of heterozygosity was mediated by mitotic crossing over (reciprocal exchange of genetic information) as well as gene conversion (non-reciprocal exchange of genetic information).