Host-derived Reactive Nitrogen Species mediate the Cryptococcus neoformans yeast-to-titan switch via fungal-derived superoxide
In the host lung, the human fungal pathogen Cryptococcus neoformans undergoes a morphological switch from small haploid yeast to large polyploid titan cell, contributing to C. neoformans virulence. Titan cells are less readily phagocytosed and can survive host nitrosative and oxidative stresses. We and others previously showed that titanization is triggered by host-relevant signals including CO2 and lung-resident bacteria, and addition of these factor is sufficient to induce titan cells in vitro. Here we investigate the molecular mechanisms that drive this transition and demonstrate that host-derived immune signals can increase the degree and frequency of titanization. Specifically, host-relevant reactive nitrogen species increase the accumulation of endogenous superoxide within cryptococcal cells, particularly within nuclei, where it can cause genotoxic stress. Consistent with this, we observe the accumulation of Rad51 protein, a marker of the double strand break repair pathway, in titanizing cultures. Blocking superoxide accumulation inhibits titanization, yet titanization also requires superoxide detoxification through Superoxide Dismutase (SOD) activity. Loss of mitochondrial Sod2 activity locks cells in the yeast phase, while Sod1 is required for the production of viable titan daughter cells. We hypothesize that the redox responsive transcription factor Yap1 in part mediates this response by regulating SOD2/SOD1. In addition, we show that Sod1 translocates to the nucleus, where it is likely involved in the detoxification of genotoxic superoxide. Together, these findings reveal a major new regulatory mechanism for the yeast-to-titan transition.