Genetic analysis of nematode cell differentiation

我们将继续研究神经元分化所需的基因， 使用线虫的六个触觉受体神经元（TRN）的功能 优雅我们之前的研究确定了这一代所需的基因， TRN的规格、维护和功能。在上一个资助期内，我们1） 确定了一个“双阴性”类型的神经元规格，其中抑制 转录因子阻止阻遏物转录因子的表达; 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 乙酰转移酶作为敏感背景，以确定神经元所需的其他基因 生长，寻找蛋白质降解和间隙连接功能所需的基因; 4） 发现了一种新的平衡系统，需要适当的神经突生长， 蛋白质降解机制与HSP 90分子伴侣系统相反; 5） 证明了百万突变项目（MMP）菌株的有用性， 来自沃特斯顿和莫尔曼实验室的完全测序菌株，作为一种工具， 基因发现和鉴定了许多新的触摸突变体; 6）研究了神经元 并发现了14个影响它的新基因，其中包括 对于机械感觉ECM、粘附复合物、轴突引导和轴突传导是重要的。 运输;和7）发现了一种合成的鞘表型，我们将利用在 未来未来研究的总体目标是利用这些发现， 了解如何控制单个神经元的分化以及如何 机械输入被感测和修改。最重要的实验 未来的工作是阐明转导复合物的分子结构， cryo-EM和发现和分析基因的损失，导致增加， 触摸灵敏度，因为这些将是触摸的负调节器。我们还打算 继续分析我们在MMMP菌株中发现的新的触摸基因， 重新分析触摸系统的重要胆固醇结合成分， （MEC-2），利用影响TRN的合成遗传关系的发现 包被以发现和表征该过程所需的额外基因。 我们工作的健康相关性来自于新基因的发现， 人类和其他哺乳动物中相似基因之间的相互作用。

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.