Candida albicans is the most common human fungal pathogen, yet the pathogenesis of C. albicans infection remains incompletely understood. We hypothesized that C. albicans has developed evolutionarily conserved mechanisms to invade disparate hosts and tested whether Toll mutant flies could serve as a model host for high-throughput screening of C. albicans virulence genes. We screened 34 C. albicans mutants defective in putative transcription factor genes (see http://www.tigr.org/tigr-scripts/e2k1/qzhao/page.cgi?num=1 ) by means of a previously established model of invasive candidiasis in Toll mutant flies. C. albicans mutants that displayed attenuated virulence in flies were subsequently tested for virulence in a mouse model of hematogenous candidiasis. Of the 34 C. albicans mutants tested, only the prototrophic cas5Delta/Delta mutant (strain VIC1186) exhibited attenuated virulence in Toll mutant flies that was restored in the complemented strain (VIC1190). Similarly, BALB/c mice infected intravenously with the cas5Delta/Delta mutant had significantly better survival and a lower fungal burden in kidneys and spleen than did those infected with the isogenic wild-type strain DAY185. CAS5 encodes a key transcriptional regulator of genes involved in cell wall integrity and lacks an orthologue in Saccharomyces cerevisiae. Our findings support the notion that Drosophila melanogaster is a promising model for large-scale studies of genes involved in the pathogenesis of C. albicans infection in mammals.