Phospholipase C (PLC) is crucial for the viability of Saccharomyces cerevisiae and Candida albicans, growth and development of filamentous fungi, and virulence of Cryptococcus neoformans (Cn). To investigate the mechanism of PLC-mediated signalling in C. neoformans, we established that, similar to the PLCδ isoform in mammalian cells, CnPlc1 hydrolyses phosphatidylinositol 4,5-bisphosphate (PIP2) to produce inositol trisphosphate (IP3). However, CnPlc1 does not contribute significantly to calcineurin activation which, in mammalian cells, is triggered by IP3-induced calcium release. Rather, CnPlc1-derived IP3 serves as a precursor for inositol polyphosphate (IP) conversion to more complex IPs (IP4-8). Using molecular and biochemical approaches, we characterized two IP kinases, CnArg1 and CnKcs1, and showed that they phosphorylate IP3 and IP6 respectively. We also showed that similar to CnPlc1, CnArg1 and CnKcs1 are essential for thermotolerance, cell wall integrity, mating, expression of virulence traits (melanization and secretion of phospholipase B) and virulence in the Galleria mellonella infection model. Our recent findings indicate that IPs derived from Arg1 and Kcs1 affect virulence-related proteins at the transcriptional and post-translational level, and the mechanism of this regulation will be discussed. In summary, our findings show that a key biochemical function of CnPlc1 is production of IP3 for synthesis of complex IPs and that the inositol polyphosphate anabolic pathway is essential for homeostasis and virulence of C. neoformans.