GENE STRUCTURE AND TRANSCRIPTIONAL REGULATION

基因结构和转录调控

• 批准号: 2174234
• 负责人: RICHARD A FIRTEL
• 金额: $ 42.23万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2174234
• 关键词: DNA; DNA binding protein; Dictyostelium; Drosophilidae; antisense nucleic acid; autoradiography; biological signal transduction; cell cycle; cell differentiation; cell type; chemical binding; chromosome complement; computer program /software; cyclic AMP receptors; cytogenetics; developmental genetics; eukaryote; fungal genetics; gel electrophoresis; gene complementation; gene expression; gene mutation; genetic library; genetic manipulation; genetic mapping; genetic promoter element; genetic regulation; genetic transcription; genetic translation; in situ hybridization; laboratory rabbit; language translation; life cycle; messenger RNA; molecular cloning; mutant; nucleic acid sequence; plant growth /development; plasmids; protein biosynthesis; radiotracer; site directed mutagenesis; structural genes

This application directs itself at further elucidating the molecular mechanisms regulating prestalk and prespore differentiation in Dictyostelium. The two cell types are spatially localized in the anterior (prestalk) and posterior (prespore) of the migrating slug. Signals of cAMP transduced through cell surface receptors activate the expression of genes unique or predominantly expressed in either one of the two cell types. Using diverse molecular and genetic approaches, we propose to study the cAMP receptor-activated signal transduction pathway. We will purify and clone the transacting factor that controls one class of induced prestalk genes and examine its putative posttranslational modification in response to cAMP receptor activation. The cis-acting sequences and trans-acting factor(s) controlling the expression of the prestalk, cell-type-specific Dd-ras genes and a coordinately regulated, cAMP-inducible class of prespore genes will be further defined. The trans-acting factor associated with a conserved regulatory element in the prespore promoters will be purified and analyzed. Using molecular complementation with genomic libraries, we will rescue existing mutants and mutations we will generate, blocked in the expression of cAMP-induced genes. With the plasmids that provided rescue, we will clone and characterize the wild-type genes. We will investigate the molecular mechanisms regulating spatial localization of the prestalk and prespore gene expression using prestalk and prespore specific promoter/beta-gal fusions. These will be used to examine the molecular basis of an anterior to posterior gradient of prespore gene expression within the prespore zone of the slug and the role of the extracellular morphogens DIF and cAMP in establishing and maintaining this gradient. We will also examine the role of the cell cycle in regulating cell-type differentiation and the ontogeny of prestalk and prespore cells. These diverse approaches will be integrated with the goal of understanding the integrated pathways that regulate cellular differentiation in this organism.

该应用将自己引向进一步阐明分子 调节预偏差和前区分化的机制 dictyostelium。两种细胞类型在空间上位于前部 迁移sl的（prestalk）和后部（前）。营地的信号 通过细胞表面受体转导的基因表达 在两种细胞类型中的任何一种中唯一或主要表达。 使用各种分子和遗传方法，我们建议研究 cAMP受体激活的信号转导途径。我们将净化并 克隆控制一类诱导Prestalk的交易因子 基因并检查其推定的翻译后修饰 训练受体激活。顺式作用序列和反式序列 控制Prestalk，细胞型特异性的表达因素 DD-RAS基因和协调的训练营前期类别 基因将进一步定义。与 前启动子中保守的监管要素将被纯化，并且 分析。使用基因组文库的分子互补，我们将 营救我们将产生的现有突变体和突变，被阻止 表达营地诱导的基因。带有提供救援的质粒， 我们将克隆并表征野生型基因。我们将调查 调节Prestalk和Prestalk空间定位的分子机制 使用Prestalk和特定于特异性的前基因表达 启动子/β-gal融合。这些将用于检查分子 前后基因前梯度的基础 在sl的前区域内和细胞外的作用 在建立和维持这一梯度方面的形态植物和扎营。我们 还将检查细胞周期在调节细胞类型中的作用 分化以及预释放细胞和前细胞的个体发育。这些 各种方法将集成，以了解 调节细胞分化的综合途径 生物。