Cytoplasmic transport of mRNAs in the myelin sheath

髓鞘中 mRNA 的细胞质转运

• 批准号: 6642458
• 负责人: DAVID R COLMAN
• 金额: $ 4.03万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6642458
• 关键词: RNA binding protein; South America; biological signal transduction; cytoplasm; cytoskeleton; density gradient ultracentrifugation; double stranded RNA; genetic translation; immunoprecipitation; interdisciplinary collaboration; intracellular transport; laboratory mouse; laboratory rat; mass spectrometry; messenger RNA; myelin; nucleic acid quantitation /detection; oligodendroglia; protein biosynthesis; protein protein interaction; ribonucleoproteins; yeast two hybrid system

DESCRIPTION (provided by applicant) This is a collaborative proposal between the laboratories of David R. Colman at Mount Sinai, NY, and of Graciela L. Boccaccio at the Institute Leloir (formerly Institute for Biochemical Research Campomar Foundation), Buenos Aires, Argentina. This proposal extend our ongoing work on the Functional Organization of the Myelin Axoglial Junction (grant # 1R01 NS40560; 07/01/00 - 06/30/05) to the study of intracellular elements of the glial cell cytoplasm adjacent to the junction. Specifically, we are aimed to elucidate the cell mechanisms mediating the localized synthesis of proteins in the myelinating cells of the central nervous system, that has being for long time of major interest to both, the applicant's and the foreign collaborator's research group. We will investigate the participation of the double stranded RNA binding protein Staufen in the localization and translation of mRNAs in oligodendrocytes. This is likely relevant to two related processes: 1. synthesis of components required for membrane maintenance and remodeling, and 2, signal transduction at the oligodendrocyte side of the axoglial junction. We will 1) use mass spectrometry to unambiguously identify novel protein components of the mRNA localization apparatus and 2) begin to asses the role of Staufen and novel components in mRNA localization. An important aspect of this proposal is its complementary nature combining the scientific interests and the methodological strength of two laboratories. Dr. Boccaccio's group have developed primary culture of oligodendrocytes and biochemical analysis of brain subcellular components, as well as a functional assay for mRNA localization in cultured cells. A number of methods that are strictly required for the present project have been established in our laboratory in Mount Sinai, namely, protein analysis via mass spectrometry and analysis of protein-protein interactions by yeast two-hybrid and "pull-down" experiments. Thus, this project will allow the training of the foreign investigator and Ph.D. students on state-of-the-art techniques that are currently not established at their home institute, thus representing a valuable contribution

描述（由申请人提供） 这是大卫R.科尔曼在西奈山，纽约州，和Graciela L。薄伽丘在研究所Leloir（原研究所生化研究Campomar基金会），布宜诺斯艾利斯，阿根廷。该提议将我们正在进行的关于髓鞘轴神经胶质连接的功能组织的工作（批准号1 R 01 NS 40560; 07/01/00 - 06/30/05）扩展到对连接附近的神经胶质细胞胞质的细胞内元件的研究。具体地说，我们的目的是阐明介导中枢神经系统髓鞘细胞中蛋白质的局部合成的细胞机制，这是申请人和外国合作者的研究小组长期以来的主要兴趣。我们将研究参与的双链RNA结合蛋白Staufen在寡突胶质细胞中的mRNA的定位和翻译。这可能与两个相关过程有关：1。合成膜维持和重塑所需的组分，和2，在轴突胶质连接的少突胶质细胞侧的信号转导。 我们将1）使用质谱法明确鉴定mRNA定位装置的新蛋白质组分，2）开始评估施陶芬和新组分在mRNA定位中的作用。 这一建议的一个重要方面是其互补性，结合了两个实验室的科学兴趣和方法优势。Boccaccio博士的小组已经开发了少突胶质细胞的原代培养和脑亚细胞成分的生化分析，以及培养细胞中mRNA定位的功能测定。本项目严格要求的一些方法已经在我们西奈山的实验室建立，即通过质谱法进行蛋白质分析，通过酵母双杂交和“下拉”实验分析蛋白质-蛋白质相互作用。因此，该项目将允许培训外国研究者和博士。学生在国家的最先进的技术，目前没有建立在他们的家乡学院，从而代表了宝贵的贡献