Vacuolar proton-translocating ATPase is required for antifungal resistance and virulence of Candida glabrata

• Published in: PloS one 2019-01, Vol.14 (1), p.e0210883-e0210883
• Main Authors: Minematsu, Asuka, Miyazaki, Taiga, Shimamura, Shintaro, Nishikawa, Hiroshi, Nakayama, Hironobu, Takazono, Takahiro, Saijo, Tomomi, Yamamoto, Kazuko, Imamura, Yoshifumi, Yanagihara, Katsunori, Kohno, Shigeru, Mukae, Hiroshi, Izumikawa, Koichi
• Format: Article
• Language: English
• Subjects: Adenosine triphosphatase; Amphotericin B; Analysis; Animals; Antifungal agents; Antifungal Agents - pharmacology; ATPases; Azoles; Biochemistry; Biology and Life Sciences; Calcineurin; Calcineurin inhibitors; Calcium; Calcium ions; Candida; Candida glabrata; Candida glabrata - drug effects; Candida glabrata - enzymology; Candida glabrata - pathogenicity; Candidiasis; Candidiasis - drug therapy; Candidiasis - microbiology; Cell growth; Clonal deletion; Drug Resistance, Fungal - genetics; Drug Resistance, Fungal - physiology; Drug Resistance, Multiple, Fungal - physiology; Drug Synergism; Environmental stress; Female; Fluconazole; Fluconazole - pharmacology; Fungal Proteins - antagonists & inhibitors; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungi; Fungicides; Gene Deletion; Genes, Fungal; Genetic aspects; Genomes; H+-transporting ATPase; Heterocyclic compounds; Homeostasis; Hospitals; Humans; Infectious diseases; Inhibitors; Iron; Laboratory animals; Macrolides - pharmacology; Manganese ions; Medicine; Medicine and Health Sciences; Membranes; Mice; Mice, Inbred BALB C; Microbial drug resistance; Multidrug resistance; pH effects; Physical Sciences; Physiological aspects; Reagents; Research and Analysis Methods; Synergistic effect; Tacrolimus; University graduates; Vacuolar Proton-Translocating ATPases - antagonists & inhibitors; Vacuolar Proton-Translocating ATPases - genetics; Vacuolar Proton-Translocating ATPases - metabolism; Vacuoles - metabolism; Virulence; Virulence (Microbiology); Virulence - drug effects; Virulence - genetics; Virulence - physiology; Voriconazole; Voriconazole - pharmacology; Yeast; Zinc
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0210883

Vacuolar proton-translocating ATPase (V-ATPase) is located in fungal vacuolar membranes. It is involved in multiple cellular processes, including the maintenance of intracellular ion homeostasis by maintaining acidic pH within the cell. The importance of V-ATPase in virulence has been demonstrated in several pathogenic fungi, including Candida albicans. However, it remains to be determined in the clinically important fungal pathogen Candida glabrata. Increasing multidrug resistance of C. glabrata is becoming a critical issue in the clinical setting. In the current study, we demonstrated that the plecomacrolide V-ATPase inhibitor bafilomycin B1 exerts a synergistic effect with azole antifungal agents, including fluconazole and voriconazole, against a C. glabrata wild-type strain. Furthermore, the deletion of the VPH2 gene encoding an assembly factor of V-ATPase was sufficient to interfere with V-ATPase function in C. glabrata, resulting in impaired pH homeostasis in the vacuole and increased sensitivity to a variety of environmental stresses, such as alkaline conditions (pH 7.4), ion stress (Na+, Ca2+, Mn2+, and Zn2+ stress), exposure to the calcineurin inhibitor FK506 and antifungal agents (azoles and amphotericin B), and iron limitation. In addition, virulence of C. glabrata Δvph2 mutant in a mouse model of disseminated candidiasis was reduced in comparison with that of the wild-type and VPH2-reconstituted strains. These findings support the notion that V-ATPase is a potential attractive target for the development of effective antifungal strategies.