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）：信号通路（例如 MAP 激酶通路）在后生动物发育过程中的许多不同组织中使用，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。该提案的长期目标是全面了解如何通过改变靶基因表达来解释来自细胞质信号（例如 MAP 激酶）的信息，从而产生特定的细胞命运。这些研究重点关注 MAP 激酶对秀丽隐杆线虫种系减数分裂进程的调节。由于 MAP 激酶介导许多物种的减数分裂进程，因此从这些研究中获得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识别作用于 MAP 激酶信号通路下游的基因，然后对关键效应器进行经典分子分析，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了解。 具体来说，DNA 微阵列将用于识别秀丽隐杆线虫种系中 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进行定义，并通过转基因分析测试 MAP 激酶反应性。然后将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这些下游效应子（无论是转录因子还是转录靶标）在体内介导MAP激酶依赖性减数分裂进程中的需求。这些实验应该共同构建对用于响应常用信号通路而产生特定细胞命运的机制的全面理解。