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The heterothallic basidiomycetous fungus  Cryptococcus neoformans  has two mating types,  MAT  a  and  MAT α. Morphological progression of bisexual reproduction in  C. neoformans  is as follows: yeast to hyphal transition, filament extension, basidium formation, meiosis, and sporulation.  C. neoformans  Cdk-related kinase 1 ( CRK1 ) is a negative regulator of bisexual mating. In this study, we characterized the morphological features of mating structures in the  crk1  mutant and determined the genetic interaction of  CRK1  in the regulatory networks of sexual differentiation. In the bilateral  crk1  mutant cross, despite shorter length of filaments than in the wild-type cross, dikaryotic filaments and other structures still remained intact during bisexual mating, but the timing of basidium formation was approximately 18 h earlier than in the cross between wild type strains. Furthermore, gene expression analyses revealed that  CRK1  modulated the expression of genes involved in the progression of hyphal elongation, basidium formation, karyogamy and meiosis. Phenotypic results showed that, although deletion of  C. neoformans CRK1  gene increased the efficiency of bisexual mating, filamentation in the  crk1  mutant was blocked by  MAT2  or  ZNF2  mutation. A bioinformatics survey predicted the  C. neoformans  GATA transcriptional factor Gat1 as a potential substrate of Crk1 kinase. Our genetic and phenotypic findings revealed that  C. neoformans   GAT1  and  CRK1  formed a regulatory circuit to negatively regulate  MAT2  to control filamentation progression and transition during bisexual mating.

Sexual Differentiation Is Coordinately Regulated by Cryptococcus neoformans CRK1 and GAT1