The Role of CRMP in Sema3 A Signaling & Axon Guidence

CRMP 在 Sema3 A 信号转导中的作用

• 批准号: 6943996
• 负责人: ERIC F SCHMIDT
• 金额: $ 4.07万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-10 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6943996
• 关键词: RNA interference; SDS polyacrylamide gel electrophoresis; biological signal transduction; chick embryo; confocal scanning microscopy; developmental neurobiology; gene expression; gene induction /repression; growth cones; histology; immunocytochemistry; neurogenesis; neuronal guidance; phosphoproteins; predoctoral investigator; protein protein interaction; protein structure function; yeast two hybrid system

DESCRIPTION (provided by applicant): Understanding how axons respond to guidance cues to navigate through the developing embryo to connect with the proper target is essential to combat developmental nervous system disorders and regeneration. Sema3A is a common axon repellent which causes growth cone collapse and axon paralysis in numerous types of neurons. To date, not much is known about the intracellular mechanisms that cause cytoskeletal rearrangements necessary for collapse. The collapsing-response-mediator-protein (CRMP) family of cytosolic phosphoproteins is required for Sema3A-induced growth cone collapse. This proposal aims to determine the mechanism of CRMP function at molecular, biochemical, and functional levels of analysis. First, the solved crystal of CRMP1 will be used to generate a series of targeted mutant proteins. These proteins will be used to map functional domains on CRMP1 surface. Second, a constitutively active (CA) mutant CRMP1 that we developed will be used to identify novel CRMP1-binding proteins. Interactions between CRMP1 and putative interacting proteins will be characterized and relevance to Sema3A signaling will be determined. Finally, gain-of-function (via overexpression of CA CRMP1) and loss-of function (RNAi) experiments will be performed to assess the function of CRMP proteins in intact chick embryos. The proposed plan of research will elucidate the mechanism of CRMP action in Sema3A signal transduction at molecular, biochemical, and functional levels of analysis.

描述（由申请人提供）：了解轴突如何响应指导线索以导航开发胚胎以与适当的靶标建立联系对于打击发育性神经系统疾病和再生至关重要。 SEMA3A是一种常见的轴突驱虫剂，可导致许多类型的神经元的生长锥塌陷和轴突瘫痪。 迄今为止，对于导致塌陷所需的细胞骨架重排的细胞内机制知之甚少。 SEMA3A诱导的生长锥塌陷所必需的胞质磷酸蛋白的崩溃反应 - 响应 - 中蛋白（CRMP）家族。 该建议旨在确定分子，生化和功能分析水平上CRMP功能的机制。 首先，CRMP1的溶液晶体将用于生成一系列靶向突变蛋白。 这些蛋白质将用于绘制CRMP1表面上的功能域。 其次，我们开发的组成型活性（CA）突变体CRMP1将用于鉴定新型的CRMP1结合蛋白。 将表征CRMP1与推定相互作用蛋白之间的相互作用，并确定与SEMA3A信号的相关性。 最后，将进行功能获得（通过CA CRMP1的过表达）和功能丧失（RNAi）实验，以评估CRMP蛋白在完整的雏鸡胚胎中的功能。 提出的研究计划将阐明在分子，生化和功能分析水平的SEMA3A信号转导中CRMP作用的机制。