SIGNAL TRANSDUCTION PATHWAYS FOR NEURONAL MIGRATION

神经元迁移的信号转导途径

• 批准号: 6645001
• 负责人: Christopher E Walsh
• 金额: $ 4.85万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6645001
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; cell migration; congenital brain disorder; developmental neurobiology; gene deletion mutation; genetic screening; immunoprecipitation; in situ hybridization; neurogenetics; northern blottings; phenotype; protein localization; protein protein interaction; protein structure function; protein tyrosine kinase; tissue /cell culture; yeast two hybrid system

DESCRIPTION: The proposal aims to identify and characterize components of the signaling pathways required for neuronal migration and cortical development. Although defects in Dbcn, mDab1, and beta LIS1 all lead to severe cortical abnormalities, little is known about the signaling pathways in which these gene products participate. Especially little is known about Dbcn, discovered earlier this year by Walsh and his colleagues. Aim 1 proposes to identify functionally important domains of the Dbcn protein by screening for additional XLIS/double cortex mutations. In aim 2 the distribution of Dbcn mRNA and protein will be determined, an essential first step to investigate the possible roles of Dbcn. Although database searches reveal few other proteins similar to Dbcn, an uncharacterized protein from brain known as KIAA0369 has an N-terminal domain that shares ~75% identity to Dbcn and a C-terminal domain that shares 97% identity with a cam kinase. Given the striking similarity between KIAA0369 and Dbcn and the possibility that they have similar (or competing) functions, the localization of KIAA0369 relative to Dbcn will be determined. The phenotypic similarity of mutants in Dbcn, mDab, and beta LIS1 suggests the hypothesis that these genes may function in the same critical signaling pathway. The possibility that these proteins interact directly with one another will be tested in aim 3 using immunoprecipitations and pair-wise two-hybrid assays and immunoprecipitation assays. Non-receptor tyrosine kinases such as Abl are likely to be involved in the pathways underlying cortical migration, and Walsh et al. have identified a putative Abl phosphorylation site in Dbcn. The predicted structure of Disabled as an adapter protein for non-receptor tyrosine kinases such as Abl also supports the hypothesis that non-receptor tyrosine kinases are important components of these pathways. Thus aim 3 will include studies to determine if Dbcn is a substrate for Abl and whether Abl interacts with mDab1 or beta LIS1. The final component of this proposal (aim 4) is to perform an extensive series of two-hybrid screens with Dbcn, mDab1, and beta LIS1 to screen for interacting proteins. Any proteins that are identified as interacting will be tested for binding in biochemical assays and would be candidate genes for lissencephaly.

产品说明：该提案旨在识别和表征神经元迁移和皮质发育所需的信号通路的组成部分。虽然Dbcn、mDab 1和beta LIS 1的缺陷都会导致严重的皮质异常，但对这些基因产物参与的信号通路知之甚少。 特别是对今年早些时候由沃尔什和他的同事发现的Dbcn所知甚少。 目的1提出通过筛选额外的XLIS/双皮质突变来鉴定Dbcn蛋白的功能重要结构域。 在目标2中，将确定Dbcn mRNA和蛋白的分布，这是研究Dbcn可能作用的必要的第一步。 虽然数据库搜索显示很少有其他类似于Dbcn的蛋白质，但来自大脑的未表征蛋白质KIAA 0369具有与Dbcn共享约75%同一性的N-末端结构域和与cam激酶共享97%同一性的C-末端结构域。 鉴于KIAA 0369和Dbcn之间惊人的相似性以及它们具有相似（或竞争）功能的可能性，将确定KIAA 0369相对于Dbcn的定位。Dbcn、mDab和β LIS 1突变体的表型相似性表明这些基因可能在相同的关键信号通路中起作用。 这些蛋白质直接相互作用的可能性将在目标3中使用免疫沉淀和成对双杂交测定和免疫沉淀测定进行测试。 非受体酪氨酸激酶如Abl可能参与皮质迁移的潜在途径，沃尔什等人已经鉴定了Dbcn中推定的Abl磷酸化位点。 残疾人作为非受体酪氨酸激酶如Abl的衔接蛋白的预测结构也支持非受体酪氨酸激酶是这些途径的重要组成部分的假设。因此，目标3将包括确定Dbcn是否是Abl的底物以及Abl是否与mDab 1或β LIS 1相互作用的研究。 该建议的最后一部分（目的4）是用Dbcn，mDab 1和beta LIS 1进行广泛的双杂交筛选，以筛选相互作用的蛋白质。 任何被鉴定为相互作用的蛋白质将在生物化学测定中进行结合测试，并且将是无脑畸形的候选基因。