MAP kinase signal specificity in the C. elegans germline

线虫种系中的 MAP 激酶信号特异性

• 批准号: 6692191
• 负责人: VALERIE J REINKE
• 金额: $ 28.78万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6692191
• 关键词: Caenorhabditis elegans; binding sites; biological signal transduction; cell cycle proteins; functional /structural genomics; gel mobility shift assay; gene deletion mutation; gene expression; genetic regulatory element; immunoprecipitation; microarray technology; mitogen activated protein kinase; molecular genetics; phenotype; polymerase chain reaction; regulatory gene; southern blotting; transcription factor

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.

描述（由申请人提供）：在整个后生动物发育过程中，许多不同的组织中使用信号通路（如MAP激酶通路）来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，信号引导下游效应子产生细胞最终命运的机制在很大程度上仍然是个谜。该提案的长期目标是获得关于如何通过改变靶基因表达来解释来自细胞质信号（如MAP激酶）的信息以产生特定细胞命运的全面知识。这些研究集中在MAP激酶调节秀丽隐杆线虫生殖系减数分裂进程。由于MAP激酶在许多物种中介导减数分裂进程，因此从这些研究中获得的结果将具有潜在的广泛适用性。通过采用功能基因组学方法来鉴定作用于MAP激酶信号通路下游的基因，然后对关键效应子进行经典分子分析，将获得对特定组织中MAP激酶响应的遗传和生化网络的全面知识。 具体地说，DNA微阵列将被用来确定候选转录靶基因下游的MAP激酶信号在C。线虫生殖系这些基因中的顺式作用调节位点将通过序列鉴定算法来定义，并通过转基因分析来测试MAP激酶反应性。然后检查编码转录因子的候选立即早期基因与靶基因中鉴定的顺式作用位点的结合。将使用RNA介导的干扰和遗传分析来评估在体内介导MAP激酶依赖性减数分裂进展中对这些下游效应物（无论是转录因子还是转录靶）的需求。总之，这些实验应该建立一个全面的了解机制，用于产生一个特定的细胞命运，以响应一个普遍使用的信号通路。