Regulation of membrane traffic between the ER and Golgi by SNARE-associated proteins, ZW10/RINT-1

SNARE 相关蛋白 ZW10/RINT-1 调节 ER 和高尔基体之间的膜交通

基本信息

批准号：
18370081
负责人：
TAGAYA Mitsuo
金额：
$ 10.9万
依托单位：
Tokyo University of Pharmacy and Life Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18370081/
关键词：
Svntaxin 18 ZW10 RINT-1 Dvnein Dvnactin Endoplasmic reticulum Microtubules Vesicular transport Sintaxin18 Syntaxin18

项目摘要

SNAREs (Soluble NSF attachment protein receptors) are involved in membrane fusion in the vesicular transport pathways in eukaryotic cells. We previously reported that syntaxin 18, a mammalian ER-localized SNARE, forms a complex with other SNAREs (BNIP1, p31, and Sec22b) and peripheral membrane proteins ZW10, and RINT-1). In the present study, we examined the function of a ZW10/RINT-1 complex in membrane traffic between the ER and the Golgi apparatus, and obtained the following results.1) Interaction between ZW10 and dynamitinProtein transport from the ER to the Golgi is mediated by the microtubule motor protein dynein-dynactin. We found that dynamitin, a subunit of the dynactin complex, and RINT-1 bind to the N-terminal region of ZW10 in a competitive manner. Evidence for the in vivo interaction of the N-terminal region of ZW10 with dynamitin was obtained by using nordihydroguaiaretic acid, a reagent that is known to stimulate dynein-dynactin-mediated processes. These results raise the possibility that ZW10 on the ER membrane is recruited to transport carriers through the interaction with dynamitin.2) Interaction of ZW10 with coatomer subunitsCOPI-coated vesicles are responsible for protein transport from the ER to the Golgi in mammalian cells. Binding experiments revealed that β,γ, and δ-COPI, weakly interact with ZW10, supporting the idea that ZW10 exists on COPI-coated vesicles and acts as a dynein-dynactin receptor.3) The NAG (Neuroblastoma amplified gene) product interacts with ZW10/RINT-1We discovered that NAG is a component of the syntaxin 18 complex. The N-terminal region of NAG interacts with the N-terminal region of p31, and the C-terminal region of NAG bind ZW10/RINT-1.

在真核细胞中的水泡转运途径中，网罗（可溶性NSF附着蛋白受体）参与膜融合。我们先前报道说，语法18（一种哺乳动物ER定位的圈圈）与其他网res（BNIP1，P31和SEC22B）以及外周膜蛋白ZW10和RINT-1形成复合物。在本研究中，我们检查了ZW10/RINT-1复合物在ER和高尔基体之间的膜交通中的功能，并获得了以下结果。1）ZW10与从ER到高尔基体介导的DynyitinProtein之间的相互作用是由微蛋白蛋白蛋白蛋白蛋白Dynein-dynactin介导的。我们发现Dynamitin是Dynactin复合物的亚基，RINT-1以竞争方式与ZW10的N末端区域结合。通过使用Nordihydroguaiarotic Acid获得了ZW10 N末端区域与ZW10的体内相互作用的证据，该试剂已知可以刺激动力蛋白 - 二奈乳蛋白介导的过程。这些结果增加了ER膜上的ZW10通过与Dynamitin的相互作用募集以运输载体。2）ZW10与座椅亚基粘膜涂层的蔬菜的相互作用是导致乳腺细胞中ER到Golgi的蛋白质转运。结合实验表明，β，γ和δ-COPI与ZW10弱相互作用，支持了ZW10存在于copi涂层蔬菜上的想法，并且充当Dynein-dynactin接收器。3）NAG（Neuroblastoma amplastoma amplastoma amplifier gene）产品与ZW10/RINT-RINT-1 WEENSTENT IS NAG相互作用。 NAG的N末端区域与p31的N末端区域相互作用，NAG的C末端区域结合ZW10/RINT-1。