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细胞内的表达和调控，以及它们与细胞质膜、细胞骨架、细胞质基质和细胞核的关系。本研究分析的靶分子和结构元件为膜小窝、连接蛋白。CD-38分子，质膜中的膜底涂层;细胞质基质中的Ca/钙调蛋白依赖性蛋白激酶IV（CaMK-IV）、二酰基甘油激酶（DGK）、蛋白激酶C（PKC）和囊泡相关小G蛋白;核输入因子和细胞核中的立即早期基因。GPI锚定蛋白的隔离，尽管小窝中的鞘糖脂和鞘磷脂是组成性的，但它们被证明是交联的结果。CD-38是一种具有环ADP-核糖合成酶活性的细胞表面抗原，定位于胶质细胞，而非神经元，提示了一种新的功能性途径。 ...更多信息 环ADP-核糖合酶在神经元信号转导中的作用。详细阐明了肌营养不良蛋白在肌细胞膜底被中的定位，并在超微结构上显示了其与细胞骨架和外质表面结构的拓扑关系。阐明了DGK、CaMK-IV和PKC在分子结构和定位上的异质性，并揭示了DGK-IV和CaMK-IV的核定位。在培养细胞中阐明了新的细胞质组分的鉴定及其在核蛋白输入中的重要性。在体内细胞核中，在各种外源性刺激（如疼痛）后，几个立即早期基因的表达发生了变化。一个新的立即早期基因被确定从海马，暗示其功能参与突触可塑性。一个小的G蛋白，拉布12被证明是参与心房肌细胞的分泌囊泡运输。总之，所有这些发现对于理解与细胞超微结构相关的信号转导机制具有极其重要的意义。少

项目成果

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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。