Ras signaling activates glycosylphosphatidylinositol (GPI) anchor biosynthesis via the GPI–N-acetylglucosaminyltransferase (GPI–GnT) in Candida albicans

Abstract

The ability of <i>Candida albicans</i> to switch between yeast to hyphal form is a property that is primarily associated with the invasion and virulence of this human pathogenic fungus. Several glycosylphosphatidylinositol (GPI)-anchored proteins are expressed only during hyphal morphogenesis. One of the major pathways that controls hyphal morphogenesis is the Ras-signaling pathway. We examine the cross-talk between GPI anchor biosynthesis and Ras signaling in <i>C. albicans.</i> We show that the first step of GPI biosynthesis is activated by Ras in <i>C. albicans</i> This is diametrically opposite to what is reported in <i>Saccharomyces cerevisiae</i> Of the two <i>C. albicans</i> Ras proteins, CaRas1 alone activates GPI-GnT activity; activity is further stimulated by constitutively activated CaRas1. CaRas1 localized to the cytoplasm or endoplasmic reticulum (ER) is sufficient for GPI-GnT activation. Of the six subunits of the GPI-<i>N</i>-acetylglucosaminyltransferase (GPI-GnT) that catalyze the first step of GPI biosynthesis, CaGpi2 is the key player involved in activating Ras signaling and hyphal morphogenesis. Activation of Ras signaling is independent of the catalytic competence of GPI-GnT. This too is unlike what is observed in <i>S. cerevisiae</i> where multiple subunits were identified as inhibiting Ras2. Fluorescence resonance energy transfer (FRET) studies indicate a specific physical interaction between CaRas1 and CaGpi2 in the ER, which would explain the ability of CaRas1 to activate GPI-GnT. CaGpi2, in turn, promotes activation of the Ras-signaling pathway and hyphal morphogenesis. The <i>Cagpi2</i> mutant is also more susceptible to macrophage-mediated killing, and macrophage cells show better survival when co-cultured with <i>Cagpi2</i>.