Vesicle formation and recycling in the secretory pathway - from yeast to higher plants

分泌途径中囊泡的形成和回收——从酵母到高等植物

基本信息

批准号：
10215210
负责人：
NAKANO Akihiko
金额：
$ 74.3万
依托单位：
RIKEN
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10215210/
关键词：
protein secretion membrane traffic ARF1 RER1 SEC12 Ara6 bulb gravitropism SAR1 RER2 ARA6 RMA1 ARE1 SAY1 GFP ARA4

项目摘要

Studies on yeast :1) We analyzed ts alleles of Arf1 GTPase and isolated multicopy suppressors. We suggested that different GEFs and GAPs are involved in an allele-specific manner. 2) We demonstrated that Rer1p is a sorting receptor in the Golgi that retrieves a subset of ER membrane proteins. We also identified the Golgi-localization signals of Rer1p and showed that they interact with COPI components. 3) We identified ER-localization signals of Sec12p and Sec71p. We also crystalized Sec12p and analyzed its 3-D structure. 4) We showed that ergosterol is essential for targeting of the tryptophan permease Tat2p to the plasma membrane. 5) We demonstrated that both Rer2p and its homologue Srt1p have the cis-prenyltransferase activity but show different characteristics.Studies on plants :1) We introduced dominant negative mutants of AtARF1 into plant cells and showed that its expression inhibits ER-Golgi traffic. 2) We identified Ara6 and Ara7 as new members of the Rab GTPase family and showed that they function in endosomes. We showed that Ara6 has uniques features distinct from other conventional Rab GTPases. 3) We showed that Arabidopsis RMA1, originally isolated as a suppressor of the yeast sec15 mutant, encodes a novel membrane-bound ubiquitin ligase. 4) We showed that vacuole membranes of Arabidopsis plants as visualized by γTIP-GFP display very dynamic and complex structures. 5) We demonstrated from the analysis of Arabidopsis shoot gravitropism mutants that vacuole biogenesis plays very important roles in the sensing of the gravity.

酵母的研究：1）我们分析了ARF1 GTPase的TS等位基因和孤立的多拷贝补充剂。我们建议以特定于等位基因的方式涉及不同的GEF和差距。 2）我们证明了RER1P是Golgi中的分类接收器，可检索ER膜蛋白的子集。我们还鉴定了RER1P的高尔基定位信号，并表明它们与COPI成分相互作用。 3）我们确定了SEC12P和SEC71P的ER定位信号。我们还将SEC12P结晶并分析了其3-D结构。 4）我们表明，麦角固醇对于将色氨酸渗透酶Tat2p靶向质膜至关重要。 5）我们证明了RER2P及其同源性SRT1P都具有顺式 - 丙二醇转移酶活性，但显示出不同的特征。研究植物：1）我们将Atarf1的显性负突变体引入了植物细胞中，并表明其表达抑制了ER-Golgi的交通。 2）我们将ARA6和ARA7确定为RAB GTPase家族的新成员，并表明它们在内体中起作用。我们表明ARA6具有与其他常规RAB GTPases不同的独特特征。 3）我们表明，拟南芥rma1最初是分离为酵母Sec15突变体的抑制剂，编码了一种新型膜结合的泛素连接酶。 4）我们表明，通过γTIP-GFP可视化的拟南芥植物的真空膜表现出非常动态和复杂的结构。 5）我们从拟南芥芽引力突变体的分析中证明了真空生物发生在重力的敏感性中起着非常重要的作用。