Cryptococcusis a fungal pathogen whose virulence relies on proliferation in and dissemination to host sites, and on synthesis of a defensive yet metabolically costly polysaccharide capsule. Regulatory pathways required forCryptococcusvirulence include a GATA-like transcription factor, Gat201, that regulates Cryptococcal virulence in both capsule-dependent and capsule-independent ways. Here we show that Gat201 is part of a negative regulatory pathway that limits fungal survival. RNA-seq analysis found strong induction ofGAT201expression within minutes of transfer to host-like media at alkaline pH. Microscopy, growth curves, and colony forming units to test viability show that in host-like media at alkaline pH wild-typeCryptococcus neoformansyeast cells produce capsule but do not bud or maintain viability, whilegat201Δcells make buds and maintain viability, yet fail to produce capsule.GAT201is required for transcriptional upregulation of a specific set of genes in host-like media, the majority of which are direct Gat201 targets. Evolutionary analysis shows that Gat201 is conserved within pathogenic fungi but lost in model yeasts. This work identifies the Gat201 pathway as controlling a trade-off between proliferation, which we showed is repressed byGAT201, and production of defensive capsule. The assays established here will allow characterisation of the mechanisms of action of the Gat201 pathway. Together, our findings urge improved understanding of the regulation of proliferation as a driver of fungal pathogenesis.