NOVEL SYSTEM FOR MONITORING AUTOPHAGY IN THE YEAST SACCHAROMYCES-CEREVISIAE
NOVEL SYSTEM FOR MONITORING AUTOPHAGY IN THE YEAST SACCHAROMYCES-CEREVISIAE
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DOI：
10.1006/bbrc.1995.1636
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发表时间：
1995-05-05
影响因子：
3.1
通讯作者：
OHSUMI, Y
中科院分区：
文献类型：
--
作者：
NODA, T;MATSUURA, A;OHSUMI, Y
在营养饥饿条件下，酿酒酵母通过非选择性自噬将胞质成分导入液泡，并通过液泡水解酶降解。我们开发了一种监测自噬的新系统，通过构建细胞，其中修饰的液泡碱性磷酸酶在细胞质中以非活性前体形式表达。在饥饿条件下，成熟形态的前体加工和磷酸酶活性逐渐出现，成熟形态位于液泡内。APG1是自噬的重要基因，破坏APG1导致加工或磷酸酶活性丧失。这些结果表明细胞质溶胶中的前体形式通过自噬转移到液泡中，并通过液泡蛋白酶转化为活性形式。因此，通过测定磷酸酶活性可以方便、准确地测定自噬。(C) 1995学术出版社，Inc。
The yeast S. cerevisiae imports cytosolic components into the vacuole non-selectively by autophagy and degrades them by vacuolar hydrolases under nutrient starvation conditions. We developed a novel system for monitoring autophagy by constructing cells in which modified vacuolar alkaline phosphatase is expressed as an inactive precursor form in the cytosol. Under starvation conditions, the processing of the precursor to the mature form and phosphatase activity appeared gradually, and the mature form was located in the vacuole. Disruption of APG1, an essential gene for autophagy, resulted in no processing or phosphatase activity. These results indicate that the precursor form in the cytosol is transferred to the vacuole by autophagy and converted to the active form by vacuolar proteinases. Thus, autophagy could be determined easily and accurately by measuring the phosphatase activity. (C) 1995 Academic Press, Inc.