Molecular Genetical Study of the Process of Transition to Stationary Phase in Yeast

酵母向固定相转变过程的分子遗传学研究

• 批准号: 07454217
• 负责人: OHSUMI Yoshinori
• 金额: $ 1.86万
• 依托单位: (National Institute for Basic Biology) Okazaki National Research Institutes (1996)The University of Tokyo (1995)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07454217/
• 关键词: Stationary Phase; Nutrient; Starvation; Autophagy; Yeast; APG genes; apg変異

It is not known the mechanism to enter stationary phase at a molecular level. We have developed a system for monitoring autophagy by prcessing of a cytosolic from of alkaline phosphatase in the vacuole. Using it we detected induction of autophagy before reaching stationary phase. We also found that under various starvation conditions yeast cells induce bulk protein degradation by autophagy. Induction of autophagy provides a good marker to analyze the process of transition to stationary phase. During these two years, we have cloned and sequenced most of the APG genes essential for induction of autophagy. APGI encodes a novel protein kinase, but we do not know the substrates or regulation of its activity yet. We found apg13 mutation is suppressed by overexpression of APGI.Apg13p is a novel hydrophilic protein. Interestingly immunoblot analysis revealed that Apg13 is heavily phosphorylated during vegetative growth, but dephoshorylated under starvation. Other APG genes are all nobel. Chacterization of these APG gene products is now undergoing.

目前尚不清楚在分子水平上进入固定相的机制。我们开发了一种通过处理液泡中碱性磷酸酶的胞质来监测自噬的系统。使用它，我们在达到稳定期之前检测到自噬的诱导。我们还发现，在各种饥饿条件下，酵母细胞通过自噬诱导大量蛋白质降解。自噬的诱导为分析向稳定期的过渡过程提供了良好的标记。在这两年里，我们已经克隆并测序了大部分诱导自噬所必需的APG基因。 APGI 编码一种新型蛋白激酶，但我们还不知道其活性的底物或调节。我们发现APGI的过度表达会抑制apg13突变。Apg13p是一种新型亲水蛋白。有趣的是，免疫印迹分析显示 Apg13 在营养生长过程中被严重磷酸化，但在饥饿条件下会去磷酸化。其他APG基因都是诺贝尔的。这些 APG 基因产物的表征正在进行中。

项目成果

Nakamura,N.et al: "Acidification of vacuoles is required for autophagic degradation in the yeast,Saccharomyces cerevisiae." J. Biochem.193 (in press). (1997)

Nakamura,N.等人：“酿酒酵母中的自噬降解需要液泡的酸化。”

Shirahama, K., Yazaki, Y., Sakano, K., Wada, Y.and Ohsumi, Y.: "Vacuolar function in the phosphate homeostasis of the yeast Saccharomyces cerevisiae." Plant Cell Physiol.37. 1090-1093 (1996)

Shirahama, K.、Yazaki, Y.、Sakano, K.、Wada, Y. 和 Ohsumi, Y.：“酿酒酵母磷酸盐稳态中的液泡功能。”

Scott, S., H-Gravink, A.Morano, K,Noda, T.and Ohsumi, Y., and Klionsky, D.: "Cytoplasm-to vacuole targeting and autophagy employ the same machinery to deliver proteins to the yeast vacuole." Proc.Natl.Acad, Sci.93. 120304-12308 (1996)

Scott, S.、H-Gravink、A.Morano, K、Noda, T. 和 Ohsumi, Y. 和 Klionsky, D.：“细胞质到液泡的靶向和自噬采用相同的机制将蛋白质递送到酵母液泡

Matsuura, A., Wada, Y., and Ohsumi, Y: "Apglp, a novel protein kinase required for the autophagic process in Saccharomyces cerevisiae" Gene. 179(in press). (1997)

Matsuura, A.、Wada, Y. 和 Ohsumi, Y：“Apglp，酿酒酵母自噬过程所需的一种新型蛋白激酶”基因。

Funakoshi,T.et al: "Analyses of APG13 gene involved in the autophagy in yeast,Saccharomyces cerevisiae." Gene. 179 (in press). (1997)

Funakoshi,T.et al：“酿酒酵母中参与自噬的 APG13 基因分析。”