Role of TFP1 in vacuolar acidification, oxidative stress and filamentous development in Candida albicans

Abstract

The vacuolar-type H+-ATPase (V-ATPase) is a multiprotein complex consisting of the V0 and V1 sectors, and is required for vacuolar acidification and virulence in the opportunistic fungal pathogen Candida albicans. In this study, we identified C. albicans Tfp1 as a putative subunit of V-ATPase, and explored its importance in multiple cellular processes. Our results revealed that Tfp1 played an essential role in vacuolar acidification and endocytic trafficking. In addition, the tfp1Δ/Δ mutant was sensitive to alkaline pH and elevated calcium concentrations, which is characteristic of loss of V-ATPase activity. The mutant also showed hypersensitivity to metal ions which might be attributed to a defect in sequestration of toxic ions to the vacuole through proton gradient produced by V-ATPase. Interestingly, deletion of TFP1 triggered endogenous oxidative stress even without exogenous oxidants. Compared with the wild-type strain, the tfp1Δ/Δ mutant showed significantly higher ROS levels and lower expression levels of redox-related genes with the addition of hydrogen peroxide (H2O2). Western blotting analysis showed that deletion of TFP1 significantly reduced the expression of Cap1 under H2O2 treatment, which contributes to the regulation of genes involved in theoxidativestressresponse. Furthermore, the tfp1Δ/Δ mutant showed significantly impaired filamentous development in hyphal induction media, and was avirulent in a mouse model of systemic candidiasis. Taken together, our results suggested that the putative V1 subunit Tfp1 is essential for vacuolar function and C. albicans pathogenesis, and provided a promising candidate for antifungal drugs.