Candida albicans White and Opaque Cells Undergo Distinct Programs of Filamentous Growth

The ability to switch between yeast and filamentous forms is central to Candida albicans biology. The yeast-hyphal transition is implicated in adherence, tissue invasion, biofilm formation, phagocyte escape, and pathogenesis. A second form of morphological plasticity in C. albicans involves epigenetic switching between white and opaque forms, and these two states exhibit marked differences in their ability to undergo filamentation. In particular, filamentous growth in white cells occurs in response to a number of environmental conditions, including serum, high temperature, neutral pH, and nutrient starvation, whereas none of these stimuli induce opaque filamentation. Significantly, however, we demonstrate that opaque cells can undergo efficient filamentation but do so in response to distinct environmental cues from those that elicit filamentous growth in white cells. Growth of opaque cells in several environments, including low phosphate medium and sorbitol medium, induced extensive filamentous growth, while white cells did not form filaments under these conditions. Furthermore, while white cell filamentation is often enhanced at elevated temperatures such as 37°C, opaque cell filamentation was optimal at 25°C and was inhibited by higher temperatures. Genetic dissection of the opaque filamentation pathway revealed overlapping regulation with the filamentous program in white cells, including key roles for the transcription factors EFG1, UME6, NRG1 and RFG1. Gene expression profiles of filamentous white and opaque cells were also compared and revealed only limited overlap between these programs, although UME6 was induced in both white and opaque cells consistent with its role as master regulator of filamentation. Taken together, these studies establish that a program of filamentation exists in opaque cells. Furthermore, this program regulates a distinct set of genes and is under different environmental controls from those operating in white cells.
Author Summary Candida albicans is the most common human fungal pathogen, capable of growing as a commensal organism or as an opportunistic pathogen. Perhaps the best-studied aspect of C. albicans biology is the transition between the single-celled yeast form and the multicellular filamentous form. This transition is necessary for virulence, as cells locked in either state are avirulent. Here, we demonstrate that the yeast-filament transition is tightly regulated by another morphological switch, the white-opaque phenotypic switch. White cells undergo filamentation in response to a wide range of established physiological cues, while opaque cells do not. We further show that opaque cells can indeed undergo filamentation, but that they do so in response to different environmental cues than those of white cells. We define the genetic regulation of filamentous growth in opaque cells, as well as the transcriptional profile of these cell types, and contrast them with the established program of filamentation in white cells. Our results reveal a close relationship between the white-opaque switch and the yeast-hyphal transition, and provide further evidence of the morphological plasticity of this pathogen. They also establish that epigenetic switching allows two fungal cell types with identical genomes to respond differently to environmental cues.