A new pH sensor localized in the Golgi apparatus of Saccharomyces cerevisiae reveals unexpected roles of Vph1p and Stv1p isoforms
A new pH sensor localized in the Golgi apparatus of Saccharomyces cerevisiae reveals unexpected roles of Vph1p and Stv1p isoforms
复制标题
DOI：
10.1038/s41598-020-58795-w
复制
发表时间：
2020-02-05
影响因子：
4.6
通讯作者：
Morsomme, Pierre
中科院分区：
文献类型：
--
作者：
Deschamps, Antoine;Colinet, Anne-Sophie;Morsomme, Pierre
分泌途径的逐渐酸化是保守的，对真核细胞非常重要，但直到现在还没有pH传感器可用于监测酿酒酵母中早期高尔基体的pH。因此，我们开发了一种基于pHluorin的传感器，用于高尔基体内腔的体内测量。通过使用这种新的工具，我们表明，顺式和中间高尔基体的pH值等于6.6-6.7在野生型细胞在指数期。正如预期的那样，V-ATP酶失活的结果在一个接近中性的高尔基体pH值。我们还发现，令人惊讶的Vph 1 p亚型的V-ATP酶是普遍的Stv 1 p的高尔基体酸化。此外，我们观察到，在细胞质pH值的变化，高尔基体的pH值保持相对稳定，主要是由于V-ATPase。最终，这种新的探针将允许更好地理解分泌途径中的酸化和pH控制所涉及的机制。
The gradual acidification of the secretory pathway is conserved and extremely important for eukaryotic cells, but until now there was no pH sensor available to monitor the pH of the early Golgi apparatus in Saccharomyces cerevisiae. Therefore, we developed a pHluorin-based sensor for in vivo measurements in the lumen of the Golgi. By using this new tool we show that the cis- and medial-Golgi pH is equal to 6.6-6.7 in wild type cells during exponential phase. As expected, V-ATPase inactivation results in a near neutral Golgi pH. We also uncover that surprisingly Vph1p isoform of the V-ATPase is prevalent to Stv1p for Golgi acidification. Additionally, we observe that during changes of the cytosolic pH, the Golgi pH is kept relatively stable, mainly thanks to the V-ATPase. Eventually, this new probe will allow to better understand the mechanisms involved in the acidification and the pH control within the secretory pathway.