The Candida albicans vacuole is required for differentiation and efficient macrophage killing.

Yeast-hypha differentiation is believed to be necessary for the normal progression of Candida albicans infections. The emergence and extension of a germ tube from a parental yeast cell are accompanied by dynamic changes in vacuole size and morphology. Although vacuolar function is required during this process, it is unclear if it is vacuolar expansion or some other vacuolar function that is important. We previously described a C. albicans vps11Delta mutant which lacked a recognizable vacuole compartment and with defects in multiple vacuolar functions. These include sensitivities to stress, reduced proteolytic activities, and severe defects in filamentation. Herein we utilize a partially functional VPS11 allele (vps11hr) to help define which vacuolar functions are required for differentiation and which influence interaction with macrophages. Mutant strains harboring this allele are not osmotically or temperature sensitive and have normal levels of secreted aspartyl protease and carboxypeptidase Y activity but have a fragmented vacuole morphology. Moreover, this mutant is defective in filamentation, suggesting that the major role the vacuole plays in yeast-hypha differentiation may relate directly to its morphology. The results of this study support the hypothesis that vacuole expansion is required during germ tube emergence. Both vps11 mutants were severely attenuated in their ability to kill a macrophage cell line. The viability of the vps11delta mutant was significantly reduced during macrophage interaction compared to that in the control strains, while the vps11hr mutant was unaffected. This implies some vacuolar functions are required for Candida survival within the macrophage, while additional vacuolar functions are required to inflict injury on the macrophage.

Regulation of dikaryon-expressed genes by FRT1 in the basidiomycete Schizophyllum commune.

The gene FRT1 has previously been shown to induce homokaryotic fruiting in transformation recipients of the basidiomycete Schizophyllum commune. In this paper, we demonstrate by gene disruption experiments that FRT1 is dispensable for dikaryotic fruiting. Nonfruiting homokaryotic FRT1 disruptant strains exhibited enhanced aerial growth of mycelia compared to wild type. Introduction of a functional FRT1 allele into the disruptant restored the wild-type colony morphology. Transcript abundance of the dikaryon-expressed SC1 and SC4 hydrophobin genes and the SC7 gene were greatly elevated in homokaryotic FRT1 disruptant strains. Growth of the disruptant strains under continuous light was found to inhibit the elevation of SC1 and SC4 transcript levels, but not of SC7 mRNA. These data suggest that the role of FRT1 in vegetatively growing homokaryons is to act as a negative regulator of dikaryon-expressed genes.