The Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba, 277-8562, Japan.
Protoplasma. 2013 Dec;250(6):1283-93. doi: 10.1007/s00709-013-0510-2. Epub 2013 May 25.
The acidic environments in the vacuole and other acidic organelles are important for many cellular processes in eukaryotic cells. In this study, we comprehensively investigated the roles of organelle acidification in vacuolar protein localisation in Saccharomyces cerevisiae. After repressing the acidification of acidic compartments by treatment with concanamycin A, a specific inhibitor of vacuolar H(+)-ATPase (V-ATPase), we examined the localisation of GFP-fused proteins that were predicted to localise in the vacuolar lumen or on the vacuolar membrane. Of the 73 proteins examined, 19 changed their localisation to the cytoplasmic region. Localisation changes were evaluated quantitatively using the image processing programme CalMorph. The delocalised proteins included vacuolar hydrolases, V-ATPase subunits, transporters and enzymes for membrane biogenesis, as well as proteins required for protein transport. These results suggest that many alterations in the localisation of vacuolar proteins occur after loss of the acidification of acidic compartments.
液泡和其他酸性细胞器中的酸性环境对于真核细胞中的许多细胞过程很重要。在这项研究中，我们全面研究了细胞器酸化在酵母液泡蛋白定位中的作用。在用特异性的液泡 H(+)-ATP 酶（V-ATPase）抑制剂康纳霉素 A 处理以抑制酸性区室酸化后，我们检查了预测定位于液泡腔或液泡膜上的 GFP 融合蛋白的定位。在所检查的 73 种蛋白质中，有 19 种改变了它们在细胞质区域的定位。使用图像处理程序 CalMorph 对定位变化进行了定量评估。去定位的蛋白质包括液泡水解酶、V-ATPase 亚基、转运蛋白和膜生物发生的酶，以及蛋白质运输所需的蛋白质。这些结果表明，酸性区室酸化丧失后，液泡蛋白定位的许多改变发生。
