RAS MEDIATED SIGNAL TRANSDUCTION IN C ELEGANS

我们的目标是了解细胞的控制和执行机制 动物发育过程中的分化和形态发生。研究 在这笔赠款的支持下，以及其他实验室的工作已经 研究表明，线虫的外阴分化是由一种RAS- 与信号转导途径极其相似的中介信号转导途径 与哺乳动物细胞信号和癌症有关的基因。然而，如何 许多常用的基因产品的活性是准确的 对特定功能的监管仍然知之甚少。此外， 发生在信号通路下游的事件，包括执行 细胞分化和形态发生的机制在很大程度上仍未被探索。在……里面 这项建议，除了继续分析规则和功能外， 对于RAS和RAF，我们将使用遗传和分子方法来鉴定和 描述一些新的基因在调节细胞周期中的功能。 信号通路和在执行外阴分化和 形态发生。具体目标是：(1)克隆和分析 两个新发现的可能是正调控基因的功能 并进行新的筛查以确定新的调节者。我们 将确定几个新分离株的遗传和分子性质 Raf突变及Raf与Ras相互作用分析 蛋白质。我们还将尝试确定RAS的RAF独立功能。 (2)开展一系列分子遗传学实验， 确定起作用的SuR-2基因的功能和调节 外阴信号转导中Ras/MAPK下游。我们会调查的 其表达模式，剖析其基因和蛋白质结构，以及 确定其监管机构和目标。(三)开展基因筛查 用于温度敏感(TS)和非TS突变体以识别新基因 以执行外阴分化和形态发生的行为。我们会 通过遗传学、微观学和分子生物学研究这些突变体所定义的基因 分子方法。细胞相互作用的分子基础研究 明确发展模式与研究发展模式密切相关 癌症和其他人类疾病。外阴发育为我们提供了一种特殊的 用于识别难以识别的因素的化验系统 并阐明它们在体内的功能。

Our goal is to understand the mechanism of control and execution of cell differentiation and morphogenesis during animal development. Research supported by this grant, as well as work from other laboratories, has shown that vulval differentiation in C. elegans is controlled by a Ras- mediated signal transduction pathway that is extremely similar to pathways that are involved in mammalian cell signaling and cancers. However, how the activities of many commonly used gene products are accurately regulated for specific functions is still poorly understood. Moreover, events occurring downstream of the signaling pathway including execution of cell differentiation and morphogenesis remain largely unexplored. In this proposal, besides continuing to analyze the regulation and function of Ras and Raf, we will use genetic and molecular methods to identify and characterize a number of new genes for their functions in regulating the signaling pathway and in executing vulval differentiation and morphogenesis. The specific aims are: (1) We will clone and analyze the function of two newly identified genes that are likely positive regulators of the pathway and carry out new screens to identify new regulators. We will determine the genetic and molecular nature of several newly isolated Raf mutations and analyze the interaction between the mutant Raf and Ras proteins. We will also try to identify Raf-independent functions of Ras. (2) We will carry out a series of molecular and genetic experiments to determine the function and regulation of the sur-2 gene which acts downstream of Ras/MAPK in vulval signal transduction. We will investigate its expression pattern, dissect its gene and protein structure, and identify its regulators and targets. (3) We will carry out genetic screens for temperature sensitive (TS) and nonTS mutants to identify new genes that act to execute vulval differentiation and morphogenesis. We will characterize genes defined by these mutants by genetic, microscopic and molecular methods. Study of the molecular basis of cell interaction in specifying developmental pattern is closely related to the study of cancers and other human disease. Vulval development provides us a specific assay system to identify factors that would be difficult to identify in mammals and to elucidate their in vivo functions.