Phenotypic consequences of LYS4 gene disruption in Candida albicans

A BLAST search of the Candida Genome Database with the Saccharomyces cerevisiae LYS4 sequence known to encode homoaconitase (HA) revealed ORFs 19.3846 and 19.11327. Both alleles of the LYS4 gene were sequentially disrupted in Candida albicans BWP17 cells using PCR-based methodology. The null lys4 mutant exhibited lysine auxotrophy in minimal medium but was able to grow in the presence of l-Lys and -aminoadipate, an intermediate of the -aminoadipate pathway, at millimolar concentrations. The presence of d-Lys and pipecolic acid did not trigger lys4 growth. The C. albicans lys4 mutant cells demonstrated diminished germination ability. However, their virulence in vivo in a murine model of disseminated neonatal candidiasis appeared identical to that of the wild-type strain. Moreover, there was no statistically significant difference in fungal burden of infected tissues between the strains. Copyright (c) 2014 John Wiley & Sons, Ltd.