Cytoplasmic transport of mRNAs in the myelin sheath

髓鞘中 mRNA 的细胞质转运

• 批准号: 6755188
• 负责人: DAVID R COLMAN
• 金额: $ 4.06万
• 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/6755188
• 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） 这是纽约州西奈山的大卫·R·科尔曼（David R.该提案将我们在髓鞘轴线连接的功能组织（授予＃1R01 NS40560; 07/01/00-06/30/05）的持续工作扩展到研究与交界处的胶质细胞细胞胞质细胞内细胞内元件的研究。具体而言，我们的目的是阐明中枢神经系统中蛋白质局部合成的细胞机制，这些细胞在中枢神经系统的髓鞘细胞中具有很长一段时间的重大关注，申请人和外国合作者的研究小组。我们将研究双链RNA结合蛋白Staufen参与少突胶质细胞中mRNA的定位和翻译。这可能与两个相关过程有关：1。膜维护和重塑所需的组件的合成，以及2个在轴突连接的寡胶质细胞侧的信号转导。 我们将1）使用质谱法明确鉴定mRNA定位设备的新型蛋白质成分，2）开始评估Staufen和新成分在mRNA定位中的作用。 该提案的一个重要方面是它的互补性，结合了两个实验室的科学利益和方法论力量。 Boccaccio博士的小组已经开发了少突胶质细胞的原发性培养物和脑亚细胞成分的生化分析，以及在培养细胞中mRNA定位的功能分析。在我们在西奈山的实验室中建立了许多严格要求的方法，即通过质谱法分析蛋白质分析，并通过酵母两杂化和“下拉”实验对蛋白质蛋白质相互作用进行分析。因此，该项目将允许对外国调查员和博士学位的培训。目前尚未在其家庭学院建立的最先进技术的学生，因此代表了宝贵的贡献