Genetic analysis of nematode cell differentiation

线虫细胞分化的遗传分析

• 批准号: 10552106
• 负责人: MARTIN CHALFIE
• 金额: $ 82.09万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-14 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552106
• 关键词: Acetyltransferase; Address; Adhesions; Affect; Axonal Transport; Binding; Biological Process; Caenorhabditis elegans; Cell Differentiation process; Cells; Cholesterol; Collection; Complex; Cryoelectron Microscopy; Development; Disease; Extracellular Matrix; Future; Gap Junctions; Generations; Genes; Genetic; Goals; Grant; Health; Heat-Shock Proteins 90; Human; Individual; Knowledge; Maintenance; Mammals; Mechanics; Molecular Chaperones; Molecular Structure; Mutation; Nematoda; Neurites; Neuronal Differentiation; Neurons; Persons; Phenotype; Process; Public Health; Research; Specific qualifier value; Structure; System; Tissues; Touch sensation; Transcription Repressor; Translating; Translational Research; Tubulin; Visit; Work; alpha Tubulin; axon guidance; beta Tubulin; experimental study; gene discovery; genetic analysis; genetic approach; insight; mutant; novel; prevent; protein degradation; receptor; tool; transcription factor

We will continue our study of genes needed for neuronal differentiation and function using the six touch receptor neurons (TRNs) of the nematode Caenorhabditis elegans. Our previous research identified genes needed for the generation, specification, maintenance, and function of the TRNs. In the last grant period, we 1) identified a “double negative” type of neuronal specification, where an inhibitory transcription factor prevents the expression of a repressor transcription factor; 2) analyzed the effects of tubulin mutations on neuronal outgrowth; 3) used the suppression of mutations affecting a TRN-specific β-tubulin and a TRN-specific α-tubulin acetyltransferase as sensitized backgrounds to identify other genes needed for neuronal outgrowth, finding genes needed for protein degradation and gap-junction function; 4) discovered a novel balancing system needed for proper neurite outgrowth in which the protein degradation machinery is opposed by the HSP90 chaperone system; 5) demonstrated the usefulness of the Million Mutation Project (MMP) strains, a collection of completely sequence strains from the Waterston and Moerman labs, as a tool for gene discovery and identified many new touch mutants; 6) investigated neuronal ensheathment and discovered 14 new genes affecting it, including genes that are important for mechanosensory ECM, adhesion complexes, axon guidance, and axonal transport; and 7) discovered a synthetic ensheathment phenotype that we will exploit in the future. The general goal of the research going forward is to exploit these findings to understand how the differentiation of individual neurons is controlled and how mechanical inputs are sensed and modified. The most important experiments for the future are the elucidation of the molecular structure of the transduction complex using cryo-EM and the discovery and analysis of genes whose loss causes an increase in touch sensitivity, since these will be negative regulators of touch. We also intend to continue analyzing the new touch genes uncovered by our work with the MMMP strains, to revisit the analysis of an important cholesterol-binding component of the touch system (MEC-2), to exploit the discovery of a synthetic genetic relationship affecting TRN ensheathment to discover and characterize additional genes needed for this process. The health relatedness of our work comes from the discovery of new genes and new interactions among genes that are similar in humans and other mammals.

我们将继续研究神经元分化所需的基因和 线虫六个触觉感受器神经元的功能 优雅女装。我们之前的研究确定了世代所需的基因， TRN的规格、维护和功能。在上一次授权期内，我们1) 发现了一种“双重否定”类型的神经元规范，其中抑制 转录因子阻止抑制转录因子的表达；2) 分析了微管蛋白突变对神经元生长的影响；3)使用 抑制影响Trn特异性β微管蛋白和Trn特异性α微管蛋白的突变 以乙酰转移酶为致敏背景来鉴定神经元所需的其他基因 生长，寻找蛋白质降解和缝隙连接功能所需的基因；4) 发现了一种新的平衡系统，需要适当的轴突生长，其中 蛋白质降解机制遭到HSP90伴侣系统的反对；5) 展示了百万突变计划(MMPs)菌株的用处，一个集合 沃特斯顿和莫尔曼实验室的菌株全序列，作为一种工具 发现并鉴定了许多新的触觉突变体；6)研究了神经元 发现了14个影响它的新基因，其中包括 对机械感觉ECM、黏附复合体、轴突引导和轴突很重要 运输；以及7)发现了一种合成的包膜表型，我们将在 未来。未来研究的总体目标是利用这些发现来 了解单个神经元的分化是如何控制的，以及如何 机械输入被感知和修改。最重要的实验是 未来将利用以下方法阐明转导复合体的分子结构 冷冻-EM及其缺失导致基因增加的发现和分析 触摸敏感度，因为这些将是触摸的负面调节器。我们还打算 继续分析我们对MMMP菌株的研究发现的新的触摸基因， 回顾触摸系统中一个重要的胆固醇结合成分的分析 (MEC-2)，利用发现影响TRN的合成遗传关系 以发现和表征这一过程所需的其他基因。 我们工作的健康相关性来自于新基因和新技术的发现 在人类和其他哺乳动物中相似的基因之间的相互作用。