The transcription factor Ofi1 is critical for white-opaque switching in natural MTLa/α isolates of Candida albicans

Candida albicans, an opportunistic fungal pathogen, is able to switch between two distinct cell types: white and opaque. While white-to-opaque switching is typically repressed by the a1/alpha 2 heterodimer in MTLa/alpha cells, it was recently reported that switching can also occur in some natural MTLa/alpha strains under certain environmental conditions. However, the regulatory program governing white-opaque switching in MTLa/alpha cells is not fully understood. Here, we collected 90 clinical isolates of C. albicans, 16 of which possess the ability to form opaque colonies. Among the known regulators implicated in white-opaque switching, only OFI1 exhibited significantly higher expression in these 16 strains compared to the reference strain SC5314. Importantly, ectopic expression of OFI1 in both clinical isolates and laboratory strains promoted switching frequency even in the absence of N-acetylglucosamine and high CO2, the optimal condition for white-to-opaque switching in MTLa/alpha strains. Deleting OFI1 resulted in a reduction in opaque-formation frequency and the stability of the opaque cell in MTLa/alpha cells. Ofi1 binds to the promoters of WOR1 and WOR3 to induce their expression, which facilitates white-to-opaque switching. Ofi1 is conserved across the CTG species. Altogether, our study reported the identification of a transcription factor Ofi1 as the critical regulator that promotes white-to-opaque switching in natural MTLa/alpha isolates of C. albicans.