YEAST PEPTIDE PHEROMONES SYNTHESIS AND MODE OF ACTION

酵母肽信息素的合成和作用方式

• 批准号: 3270736
• 负责人: Jeremy W. Thorner
• 金额: $ 15.47万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-01-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3270736
• 关键词: Saccharomyces; adenylate cyclase; affinity chromatography; antibody formation; cell adhesion; cell cell interaction; cell cycle; cell fusion; cell membrane; eukaryote; fungal genetics; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; high performance liquid chromatography; membrane activity; messenger RNA; microorganism conjugation; molecular cloning; oligopeptides; pheromone; phosphorylation; radioimmunoassay; structural genes; temperature sensitive mutant

The two haploid cell types (a and Alpha) of the yeast Saccharomyces cerevisiae each secrete specific peptide hormone-like molecules which trigger events leading to fusion ("mating") of the cells, a relatively simple developmental sequence. Because incisive genetic and biochemical studies can be performed in yeast cells, particular those utilizing recombinant DNA, it may be possible to learn the precise molecular details of peptide hormone biosynthesis and mode of action by examining the yeast mating pheromones (a-factor and Alpha-factor). The approaches to be taken include: (1) in vitro alterations of the control regions of the cloned Alpha-factor and a-factor genes, and novel gene fusions to indicator enzymes, to dissect the cell type-specific regulation of transcription of these genes; (2) specific antibodies, inhibitors, and mutations, in conjunction with the cloned genes, to provide probes and substrates for analyzing the synthesis, membrane translocation, post-translational processing, and secretory transport of the pheromone precursor proteins (prepro-Alpha-factor and prepro-a-factor); (3) [125I]-labelled pheromones, covalent cross-linking of reactive pheromone derivatives, affinity chromatography using matrix-bound pheromones, putative genes for Alpha-factor receptor (STE2) and adenylate cyclase (CDC35), and the cloned STE5 gene, to identify pheromone receptors and to examine the mechanism of pheromonal modulation of cyclic AMP synthesis; and, (4) transcript mapping, gene disruption, and DNA sequencing, to characterize the structure and function of cloned genes whose expression is under pheromonal control. In multicellular organisms, peptide hormones are responsible for cell-cell communications that are required during embryonic development and in the coordination of life functions in adults. The under- or over-production of particular peptide hormones, or the inability to sense their presence, have been correlated with numerous human diseases (e.g. dwarfism, Cushing's disease, and diabetes). Understanding of the fundamental aspects of the synthesis and release of peptide hormones, and the regulation of these processes, as well as the biochemical bases of their mechanisms of action, is necessary for effective diagnosis and therapy of such diseases. In addition, knowledge of the biosynthesis of yeast pheromones may allow the production of medically-valuable human hormones by yeast cells on a commercial scale.

酵母菌的两种单倍体细胞类型（A和α） 酿酒酵母每个分泌特异性肽激素样分子，这些分子 触发事件导致细胞的融合（“交配”），一个相对 简单的发展序列。 因为敏锐的遗传和生化 可以在酵母细胞中进行研究，尤其是利用的细胞 重组DNA，可能可以学习精确的分子细节 通过检查酵母菌的肽激素生物合成和作用方式 交配信息素（A因子和α因素）。 要采取的方法 包括：（1）克隆的控制区域的体外改变 α因子和A因子基因以及指示剂的新基因融合 酶，剖析细胞类型的特定于转录的调节 这些基因； （2）特定抗体，抑制剂和突变 与克隆基因的结合，提供探针和底物 分析合成，膜易位，翻译后 信息素前体蛋白的加工和分泌运输 （前α因素和prepro-a因子）； （3）[125i]标签的信息素， 反应性信息素衍生物的共价交联，亲和力 使用基质结合信息素的色谱法，推定的基因 α-因子受体（Ste2）和腺苷酸环化酶（CDC35），并克隆 Ste5基因，鉴定信息素受体并检查 周期性AMP合成的信息符号调节； （4）成绩单映射， 基因破坏和DNA测序，以表征结构和 克隆基因的功能，其表达受到信息符号对照。 在多细胞生物中，肽激素负责细胞细胞 在胚胎开发和 成人生活功能的协调。 不足或过量的 特定的肽激素，或无法感知其存在的能力 与许多人类疾病相关（例如矮人，库欣 疾病和糖尿病）。 理解的基本方面 肽激素的合成和释放，这些调节 过程以及其作用机理的生化基础， 对于此类疾病的有效诊断和治疗是必要的。 在 此外，了解酵母信息素的生物合成可能允许 通过酵母细胞在A上产生医学可吸收的人类激素 商业规模。