The calcineurin pathway controls unisexual reproduction, stress responses, and virulence in C. deneoformans. Calcineurin Cna1/Cnb1 functions as the central hub in the calcineurin pathway. During unisexual reproduction, calcineurin regulates the yeast–hyphal morphological transition through Cbp1 and Cts1, and via a pathway in which Cbp1 synergistically interacts with Pbp1, whereas Cts1 inhibits the pheromone response pathway. Crz1 and Puf4 are required for sporulation. In response to different environmental stresses, Crz1 synergistically interacts with other calcineurin downstream targets via complex branched pathways. Finally, Pbp1 and Tif3 are key virulence factors downstream of calcineurin in C. neoformans.