Role of DNA Mismatch Repair and Double-Strand Break Repair in Genome Stability and Antifungal Drug Resistance in Candida albicans

Abstract

ABSTRACT Drug resistance has become a major problem in the treatment of Candida albicans infections. Genome changes, such as aneuploidy, translocations, loss of heterozygosity, or point mutations, are often observed in clinical isolates that have become resistant to antifungal drugs. To determine whether these types of alterations result when DNA repair pathways are eliminated, we constructed yeast strains bearing deletions in six genes involved in mismatch repair ( MSH2 and PMS1 ) or double-strand break repair ( MRE11, RAD50, RAD52 , and YKU80 ). We show that the mre11 Δ/ mre11 Δ, rad50 Δ/ rad50 Δ, and rad52 Δ/ rad52 Δ mutants are slow growing and exhibit a wrinkly colony phenotype and that cultures of these mutants contain abundant elongated pseudohypha-like cells. These same mutants are susceptible to hydrogen peroxide, tetrabutyl hydrogen peroxide, UV radiation, camptothecin, ethylmethane sulfonate, and methylmethane sulfonate. The msh2 Δ/ msh2 Δ, pms1 Δ/ pms1 Δ, and yku80 Δ/ yku80 Δ mutants exhibit none of these phenotypes. We observed an increase in genome instability in mre11 Δ/ mre11 Δ and rad50 Δ/ rad50 Δ mutants by using a GAL1 / URA3 marker system to monitor the integrity of chromosome 1. We investigated the acquisition of drug resistance in the DNA repair mutants and found that deletion of mre11 Δ/ mre11 Δ, rad50 Δ/ rad50 Δ, or rad52 Δ/ rad52 Δ leads to an increased susceptibility to fluconazole. Interestingly, we also observed an elevated frequency of appearance of drug-resistant colonies for both msh2 Δ/ msh2 Δ and pms1 Δ/ pms1 Δ (MMR mutants) and rad50 Δ/ rad50 Δ (DSBR mutant). Our data demonstrate that defects in double-strand break repair lead to an increase in genome instability, while drug resistance arises more rapidly in C. albicans strains lacking mismatch repair proteins or proteins central to double-strand break repair.