Molecular biological and morphological analysis of signal transduction mechanism from membrane to nuchreos

从膜到核的信号转导机制的分子生物学和形态学分析

• 批准号: 07307027
• 负责人: KONDO Hisatake
• 金额: $ 2.75万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-07307027/
• 关键词: membranous caveolae; CD-38; membranous undercoat; nuclear import factor; immediate early gene; diacylglycerol kinase; CaM kinase IV; protein kinase C; コネクシン; CD-38; カベオラ; プロティンキナーゼC; カルモジュリンキナーゼIV; 核膜孔ターゲッチング複合体; gap-junction; 細胞骨格; プロティンキ

The expression and regulation of signal-related molecules were analyzed at levels of genes and proteins in cells in-vitro and in-vivo with regard to their relation to the plasma wembrane, cytoskeletons, cytoplasmic matrix and nucleus. Targeted molecules and structural elements analyzed in this study were the membranous caveolae, connexins. CD-38 molecule, membranous undercoats in the plasma membrane ; Ca/calmodulin-dependent protein kinase IV (CaMK-IV), diacylglycerol kinase (DGK), protein kinase C (PKC) and vesicle-associated small G-proteinin in the cytoplasmic matrix ; nuclear import factors and immediate early genes in the nucleus.The sequestration of GPI-anchored proteins, glycosphingolipids and sphingomyelin in the caveolae were shown to be results of cross-linking, although the localization of caveolin there were constitutive. CD-38, a cell surface-antigen having an activity of cyclic ADP-ribose synthase, was localized in glia, but not neurons, suggesting a new way of functional … More exertion of cyclic ADP-ribose synthase in the neuroanl signal transduction. The localization of dystrophin in the membranous undercoat of muscle cells were clarified in details and its topographical relation to cytoskeletons and exoplasmic surface structures were shown in ultrastructure. The heterogeneity in molecular structure and localization was clarified for DGK,CaMK-IV and PKC,and the nuclear localization of DGK-IV and CaMK-IV was revealed. The identification of novel cytoplasmic components and their importance in the nuclear protein-import were clarified in cultured cells. In the nucleus in vivo, changes in the expression of several immediate early genes occurred after various exogenous stimuli such as pain. A novel immediately early gene was identified from the hippocampus, implicating its functional involvement in synaptic plasticity. A small G-protein, rab 12 is shown to be involved in the secretory vesicle transport in atrial muscle cells. In conclusion, all these findings are of extreme importance in understanding the signal transduction mechanism in relation to cellular ultrastructure. Less

从基因和蛋白质水平分析了信号相关分子在体外和体内细胞中的表达及其与细胞膜、细胞骨架、胞质基质和细胞核的关系。本研究分析的靶向分子和结构元件为膜小窝、连接蛋白。胞质基质中的钙/钙调蛋白依赖性蛋白激酶IV(CaMK-IV)、二酰甘油蛋白激酶(DGK)、蛋白激酶C(PKC)和囊泡相关的小G蛋白；核输入因子和即刻早期基因；GPI锚定蛋白、糖鞘磷脂和鞘磷脂在小窝中的滞留是交联性的结果，尽管小窝蛋白的定位是结构性的。CD-38是一种具有环腺苷二磷酸-核糖合成酶活性的细胞表面抗原，定位于胶质细胞，而不是神经元，提示了一种新的功能…的途径环腺苷二磷酸核糖合成酶在神经细胞信号转导中的更多作用。肌营养不良蛋白在肌细胞膜性底毛中的定位得到了详细的阐明，并在超微结构中显示了其与细胞骨架和外质表面结构的地形性关系。明确了DGK、CaMK-IV和PKC在分子结构和定位上的异质性，揭示了DGK-IV和CaMK-IV的核定位。在培养细胞中，新的细胞质成分的鉴定及其在核蛋白输入中的重要性被阐明。在体内的细胞核中，疼痛等各种外源性刺激后，几个即刻早期基因的表达发生了变化。从海马体中发现了一个新的即刻早期基因，表明它参与了突触的可塑性。Rab 12是一种小G蛋白，参与了心房肌细胞分泌囊泡的运输。综上所述，所有这些发现对于理解与细胞超微结构有关的信号转导机制具有极其重要的意义。较少

项目成果

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M. Imamoto 等人和 Y. Yoneda：“核孔靶向复合物的 58 kDa 成分参与核蛋白输入的体内证据。” EMBO J.14 (1995)。

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S.Kano および H,Ishikawa: "Localization of 47KD antigenic polypeptides in Mararia parasites by confocal laser scanning microscopy" Bio images. 3. 13-17 (1995)

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Iida,H.,Wang,L.,Nishii,K.Ookura,K,Shibata,Y.: "Identification of rab 12 as a secretory granule-associated small G-protein in atital myocytes" Circ.Res.78. 343-347 (1996)

Iida,H.,Wang,L.,Nishii,K.Ookura,K,Shibata,Y.：“鉴定 rab 12 作为初始肌细胞中分泌颗粒相关的小 G 蛋白”Circ.Res.78。