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Cryptococcus neoformans  survives host temperature and regulates cell wall integrity via a calcium-dependent phosphatase, calcineurin. However, downstream effectors of  C. neoformans  calcineurin are largely unknown. In  S. cerevisiae  and other fungal species, a calcineurin-dependent transcription factor Crz1, translocates to nuclei upon activation and triggers expression of target genes. We now show that the  C. neoformans  Crz1 ortholog (Crz1/Sp1), previously identified as a protein kinase C target during starvation, is a  bona fide  target of calcineurin under non-starvation conditions, during cell wall stress and growth at high temperature. Both the calcineurin-defective mutant, Δ cna1,  and a  CRZ1/SP1  mutant (Δ crz1 ) were susceptible to cell wall perturbing agents. Furthermore, expression of the chitin synthase encoding gene,  CHS6,  was reduced in both mutants. We tracked the subcellular localization of Crz1-GFP in WT  C. neoformans  and  Δcna1  in response to different stimuli, in the presence and absence of the calcineurin inhibitor, FK506. Exposure to elevated temperature (30–37°C vs 25°C) and extracellular calcium caused calcineurin-dependent nuclear accumulation of Crz1-GFP. Unexpectedly, 1M salt and heat shock triggered calcineurin-independent Crz1-GFP sequestration within cytosolic and nuclear puncta. To our knowledge, punctate cytosolic distribution, as opposed to nuclear targeting, is a unique feature of  C. neoformans  Crz1. We conclude that Crz1 is selectively activated by calcium/calcineurin-dependent and independent signals depending on the environmental conditions.

The Crz1/Sp1 Transcription Factor of Cryptococcus neoformans Is Activated by Calcineurin and Regulates Cell Wall Integrity