1887

Abstract

The phosphatidylcholine-specific phospholipase D1 (PLD1) in is involved in vesicle transport and is essential for sporulation. The gene encoding the homologous phospholipase D1 from () was used to study the role of PLD1 in this pathogenic fungus. and expression studies using Northern blots and reverse transcriptase-PCR showed low mRNA levels in defined media supporting yeast growth and during experimental infection, while enhanced levels of transcripts were detected during the yeast to hyphal transition. To study the relevance of during yeast and hyphal growth, an essential part of the gene was deleted in both alleles of two isogenic strains. PLD1 activity assays showed that mutants produced no detectable levels of phosphatidic acid, the hydrolytic product of PLD1 activity, and strongly reduced levels of diacylglycerol, the product of lipid phosphate phosphohydrolase, suggesting no or a negligible background PLD1 activity in the mutants. The mutants showed no growth differences compared to the parental wild-type in liquid complex and minimal media, independent of the growth temperature. In addition, growth rates of mutants in media with protein as the sole source of nitrogen were similar to growth rates of the wild-type, indicating that secretion of proteinases was not reduced. Chlamydospore formation was normal in mutants. When germ tube formation was induced in liquid media, mutants showed similar rates of yeast to hyphal transition compared to the wild-type. However, no hyphae formation was observed on solid Spider medium, and cell growth on cornmeal/Tween 80 medium indicated aberrant morphogenesis. In addition, mutants growing on solid media had an attenuated ability to invade the agar. In a model of oral candidosis, mutants showed no attenuation of virulence. In contrast, the mutant was less virulent in two different mouse models. These data suggest that is not essential for growth and oral infections. However, they also suggest that a prominent part of the phosphatidic acid and diacylglycerol pools is produced by PLD1 and that the level of these components is important for morphological transitions under certain conditions in .

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2001-04-01
2020-09-27
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