MAP kinase signal specificity in the C. elegans germline

描述(由申请人提供)：在后生动物发育过程中，许多不同的组织都使用信号通路，如MAP激酶通路，以控制重要的细胞命运决定。细胞对信号的误解可能导致癌症或发育缺陷。然而，信号引导下游效应器产生细胞最终命运的机制在很大程度上仍然是个谜。这一提议的长期目标是全面了解来自细胞质信号的信息如何通过改变目标基因的表达来解释来自细胞质信号的信息，从而产生特定的细胞命运。这些研究集中在MAP激酶对秀丽线虫生殖系减数分裂进程的调控。由于MAP激酶在许多物种中介导减数分裂进程，因此这些研究的结果可能具有广泛的适用性。通过采用功能基因组学方法来确定MAP激酶信号通路下游作用的基因，然后对关键效应进行经典的分子分析，将获得对特定组织中MAP激酶反应的遗传和生化网络的全面了解。 具体地说，DNA微阵列将被用来识别线虫生殖系MAP激酶信号下游的候选转录目标基因。这些基因中的顺式作用调控位点将通过序列识别算法确定，并通过转基因分析测试MAP激酶的响应性。然后，将检查编码转录因子的候选即刻早期基因是否与目标基因中确定的顺式作用位点结合。这些下游效应物，无论是转录因子还是转录靶标，在体内介导MAPK依赖的减数分裂进程中的需求将通过RNA介导的干扰和遗传分析来评估。总之，这些实验应该有助于全面理解用来产生特定细胞命运的机制，以响应通常使用的信号通路。

DESCRIPTION (provided by applicant): Signaling pathways such as the MAP kinase pathway are used in many different tissues throughout metazoan development to control important cell fate decisions. Misinterpretation of a signal by a cell can lead to cancer or developmental defects. However, the mechanisms by which a signal directs downstream effectors to generate the ultimate fate of the cell remain largely mysterious. The long-term objective of this proposal is to gain a comprehensive knowledge of how the information from a cytoplasmic signal such as MAP kinase is interpreted through altered target gene expression to produce a specific cell fate. These studies focus on MAP kinase regulation of meiotic progression in the Caenorhabditis elegans germ line. Because MAP kinase mediates meiotic progression in many species, the results obtained from these studies will potentially be broadly applicable. By taking functional genomics approaches to identify genes acting downstream of the MAP kinase signaling pathway, followed by classical molecular analysis of key effeetors, comprehensive knowledge of the genetic and biochemical network responding to MAP kinase in a particular tissue will be attained. Specifically, DNA microarrays will be used to identify candidate transcriptional target genes downstream of MAP kinase signaling in the C. elegans germ line. The cis-acting regulatory sites in those genes will be defined through sequence identification algorithms and tested for MAP kinase responsiveness through transgenic analysis. Candidate immediate-early genes encoding transcription factors will then be examined for binding to the cis-acting sites identified in the target genes. The requirement for these downstream effectors, whether transcription factor or transcription target, in mediating MAP kinase dependent meiotic progression in vivo will be assessed using RNA-mediated interference and genetic analysis. Together these experiments should build toward a comprehensive understanding of the mechanisms used to produce a specific cell fate in response to a generally used signaling pathway.