ANALYSIS FOR THE REGULATORY MECHANISM OF THE VESICULAR TRANSPORT BY COILED-COIL PROTEINS

GM 130及其前受体的本地化机制，GRASP 65对戈尔基装置进行了调查。（1）通过脉冲追逐亚细胞分段实验使用<35>^S-methionin，它被揭示为GM 130 amd GRASP 65本地化到Golgi设备不久后的合成。（2）形态学分析揭示了新合成的GM 130和GRASP 65在戈尔基设备的运输形式下定位为戈尔基设备的条件下，被突变Sarlp的微型注射所抑制。（3）体外translated GM130 and GRASP65 specifically bound to the purified·Golgi membrane。这些结果表明GM 130和GRASP 65定位到Golgi设备直接没有初始目标到Golgi设备，GM 130和GRASP 65是支持Golgi设备的极化的候选结构蛋白。The genes for Yiplp family transmembrane proteins which are candidates for the determinant of the localization of GM130 and GRASP65 were identified from genome data base。其中一个家族成员在Golgi设备上本地化，并通过表达取消Golgi设备。“戈尔基解散的分子机制现在正在进行调查。”戈尔基装置在接受低pH值介质的细胞治疗后也被拆除。这种治疗对GM 130和GRASP 65的影响目前正在被调查。

The localization mechanisms of GM130 and its presumed receptor, GRASP65 to the Golgi apparatus were investigated. (1) By pulse-chase subcellular fractionation experiments using <35>^S-methionin, it was revealed that GM130 amd GRASP65 localize to the Golgi apparatus soon after the synthesis. (2) Morphological analysis revealed that newly synthesized GM130 and GRASP65 were localized to the Golgi apparatus under the condition the transport form the ER to the Golgi apparatus was inhibited by the microinjection of mutant Sarlp. (3) In vitro translated GM130 and GRASP65 specifically bound to the purified・Golgi membrane. These results indicated that GM130 and GRASP65 localize to the Golgi apparatus directly without initial targeting to the Golgi apparatus suggesting that GM130 and GRASP65 are the candidate structural protein that support the polalization of the Golgi apparatus. The genes for Yiplp family transmembrane proteins which are candidates for the determinant of the localization of GM130 and GRASP65 were identified from genome data base. One of the family member proteins localized at the Golgi apparatus and its over expression disassembled the Golgi apparatus. The molecular mechanism for the Golgi disassembly is now under investigation. The Golgi apparatus is also disassembled after the treatment of cells with low pH medium. The effect of this treatment for GM130 and GRASP65 are currently investigated.