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: 6574542
• 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/6574542
• 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; 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）：信号通路（例如MAP激酶途径）在整个后生动物发育的许多不同组织中都使用，以控制重要的细胞命运决策。细胞对信号的误解会导致癌症或发育缺陷。但是，信号指导下游效应子产生细胞的最终命运的机制仍然很大程度上是神秘的。该提案的长期目标是获取有关如何通过改变靶基因表达来产生特定细胞命运的细胞质信号（例如MAP激酶）的信息。这些研究集中于秀丽隐杆线虫种系中减数分裂进程的MAP激酶调节。由于MAP激酶介导了许多物种的减数分裂进程，因此从这些研究获得的结果可能是广泛适用的。通过采用功能性基因组学方法来鉴定MAP激酶信号传导途径下游作用的基因，然后进行关键效能器的经典分子分析，将获得对遗传和生化网络对特定组织中MAP激酶响应的全面知识。 具体而言，DNA微阵列将用于识别秀丽隐杆线虫生殖系中MAP激酶信号下游的候选转录靶基因。这些基因中的顺式作用调节位点将通过序列识别算法定义，并通过转基因分析测试了MAP激酶反应性。然后，将检查编码转录因子的候选基因，然后检查与目标基因中鉴定的顺式作用位点结合。这些下游效应子（无论是转录因子还是转录靶标）在介导MAP激酶依赖性减数分裂进程中的需求将使用RNA介导的干扰和遗传分析评估。这些实验应共同建立在响应一般使用的信号传导途径的响应中，对产生特定细胞命运的机制的全面理解。