The vacuolar-type H⁺-ATPase (V-ATPase) is a highly conserved protein complex among the eukaryotic cells. We previously revealed that both the V-ATPase and the transient receptor potential (TRP) channel Yvc1 are involved in oxidative stress response (OSR). However, the relationship between V-ATPase and Yvc1 during OSR remains unknown. In this study, disruption of the V-ATPase-encoding genes VPH2 and TFP1, similar with disruption of YVC1, caused H₂O₂ hypersensitivity and enhancement of vacuolar membrane permeability (VMP) under oxidative stress. Further investigations showed that unlike the wild type strain with vacuole membrane-localized Yvc1, both vph2Δ/Δ and tfp1Δ/Δ had Yvc1 localization in the vacuole cavity, indicating that disruption of VPH2 or TFP1 impaired normal vacuolar membrane-localization of Yvc1. Interestingly, addition of CaCl₂ alleviated the growth defect of vph2Δ/Δ and tfp1Δ/Δ under oxidative stress, leading to prevention of VMP, decrease in ROS levels and activation of OSR. In contrast, addition of the Ca²⁺ chelating agent glycol-bis-(2-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA) aggravated H₂O₂ hypersensitivity of the mutants. These results showed that the V-ATPase plays an important role in maintenance of normal Yvc1 localization, which contributes to Ca²⁺ transport from the vacuoles to the cytosol for activation of OSR. This work sheds a novel light on the interaction between V-ATPase and Ca²⁺ transport for regulation of OSR in C. albicans.

液泡型H ⁺ -ATPase（V-ATPase）是真核细胞中高度保守的蛋白质复合物。我们先前发现，V-ATPase和瞬时受体电位（TRP）通道Yvc1都参与了氧化应激反应（OSR）。但是，OSR过程中V-ATPase和Yvc1之间的关系仍然未知。在这项研究中，V-ATPase编码基因VPH2和TFP1的破坏与YVC1的破坏相似，导致H ₂ O ₂超敏性和氧化应激下液泡膜通透性（VMP）的增强。进一步的研究表明，与带有液泡膜定位Yvc1的野生型菌株不同，两个vph2Δ/Δ和tfp1Δ /Δ在液泡腔中具有Yvc1定位，表明VPH2或TFP1的破坏损害了Yvc1的正常液泡膜定位。有趣的是，CaCl _2的添加减轻了氧化应激下vph2Δ /Δ和tfp1Δ /Δ的生长缺陷，从而防止了VMP，ROS水平降低和OSR活化。相反，添加Ca ²⁺螯合剂二醇-双-（2-氨基乙基醚）-N，N，N'，N'-四乙酸（EGTA）会加重H ₂ O ₂突变体的超敏性。这些结果表明，V-ATPase在维持正常Yvc1定位中起着重要作用，这有助于Ca ²⁺从液泡向细胞质的转运，从而激活OSR。这项工作为白念珠菌中OSR的调控提供了V-ATPase和Ca ²⁺转运之间相互作用的新思路。