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Thevirulence ofCandida albicans mutants lacking one or bothcopies ofRAD52 , a gene involved in homologous recombination(HR), was evaluated in a murine model of hematogenously disseminatedcandidiasis. In this study, the virulence of therad52 Δ mutant was dependent upon the inoculumconcentration. Mice survived at a cell inoculum of 1 ×106 , but there was a decrease in survival time at dosages of1.5 × 106 and especially at 3 ×106 cells per animal. The heterozygoteRAD52/rad52 behaved like wild type, whereas a reintegrant strain was intermediatein its ability to cause death compared to these strains and to theavirulentrad52/rad52 null at inocula of 1 ×106 and 1.5 × 106 cells. A double mutant,lig4/lig4/rad52/rad52 , was avirulent at all inocula used. PCRanalysis of theRAD52 and/orLIG4 loci showed thatall strains recovered from animals matched the genotype of theinoculated strains. Analysis of the electrophoretical karyotypesindicated that the inoculated, reintegrant strain carried a largedeletion in one copy of chromosome 6 (the shortest homologue, orChr6b). Interestingly, truncated Chr6b was regenerated in all thestrains recovered from moribund animals using the homologue as atemplate. Further, regeneration of Chr6b was paralleled by an increasein virulence that was still lower than that of wild type, likelybecause of the persistent loss of heterozygosity in the regeneratedregion. Overall, our results indicate that systemic candidiasis candevelop in the absence of HR, but simultaneous elimination of bothrecombination pathways, HR and nonhomologous end-joining, suppressesvirulence even at very highinocula.

infection and immunity

Virulence and Karyotype Analyses of rad52 Mutants of Candida albicans : Regeneration of a TruncatedChromosome of a Reintegrant Strain ( rad52 / RAD52 ) in the Host