Proline catabolism is a key factor facilitating Candida albicans pathogenicity.

Candida albicans, the primary etiology of human mycoses, is well-adapted to catabolize proline to obtain energy to initiate morphological switching (yeast to hyphal) and for growth. We report that put1-/- and put2-/- strains, carrying defective Proline UTilization genes, display remarkable proline sensitivity with put2-/- mutants being hypersensitive due to the accumulation of the toxic intermediate pyrroline-5-carboxylate (P5C), which inhibits mitochondrial respiration. The put1-/- and put2-/- mutations attenuate virulence in Drosophila and murine candidemia models and decrease survival in human neutrophils and whole blood. Using intravital 2-photon microscopy and label-free non-linear imaging, we visualized the initial stages of C. albicans cells infecting a kidney in real-time, directly deep in the tissue of a living mouse, and observed morphological switching of wildtype but not of put2-/- cells. Multiple members of the Candida species complex, including C. auris, are capable of using proline as a sole energy source. Our results indicate that a tailored proline metabolic network tuned to the mammalian host environment is a key feature of opportunistic fungal pathogens. Author summary: Candida albicans is listed as one of four "critical priority" fungal pathogens by the World Health Organization (WHO). Intriguingly, C. albicans is a natural commensal organism thriving in symbiosis with other components of the human microflora. Given the opportunistic character of C. albicans, it is important to understand how nutrient availability within host microenvironments contribute to the transition from commensal to pathogenic growth. Here, we report that proline is catabolized by C. albicans as an important energy source, a characteristic that is conserved among other pathogenic Candida species, including the multidrug resistant C. glabrata and C. auris. Using different infection models and applying a state-of-the-art intravital imaging technique to visualize C. albicans cells infecting kidneys in a living mouse, we observed that strains unable to utilize proline exhibit significantly reduced virulence and filamentous hyphal growth. Genetic dissection of the Proline UTilization (PUT) pathway and deciphering the control mechanisms governing proline use led to the discovery that proline is toxic to cells unable to catabolize this amino acid. Our findings provide novel insights implicating proline metabolism as a key determinant of pathogenic fungal growth. [ABSTRACT FROM AUTHOR]