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: 3270738
• 负责人: Jeremy W. Thorner
• 金额: $ 17.23万
• 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/3270738
• 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 Alpha） 每种酿酒酵母都分泌特定的肽激素样分子， 触发事件导致细胞融合（“交配”），相对而言 简单的发展顺序。 因为精辟的遗传和生化 研究可以在酵母细胞中进行，特别是那些利用 重组DNA，或许可以了解精确的分子细节 通过检查酵母来研究肽激素的生物合成和作用方式 交配信息素（a 因子和 Alpha 因子）。 应采取的方法 包括：（1）克隆体控制区的体外改变 α-因子和a-因子基因，以及新的基因融合来指示 酶，剖析细胞类型特异性转录调节 这些基因； (2) 特异性抗体、抑制剂和突变， 与克隆基因结合，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 （前阿尔法因子和前α因子）； (3) [125I]-标记的信息素， 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，假定的基因 α因子受体（STE2）和腺苷酸环化酶（CDC35），以及克隆 STE5基因，识别信息素受体并检查其机制 环 AMP 合成的信息素调节；以及，（4）转录本映射， 基因破坏和 DNA 测序，以表征结构和 其表达受信息素控制的克隆基因的功能。 在多细胞生物中，肽激素负责细胞间的相互作用。 胚胎发育和发育过程中所需的通讯 成人生活功能的协调。 生产不足或过剩 特定的肽激素，或无法感觉到它们的存在，已经 与许多人类疾病（例如侏儒症、库欣病 疾病和糖尿病）。 对基本方面的理解 肽类激素的合成和释放及其调节 过程及其作用机制的生化基础， 对于此类疾病的有效诊断和治疗是必要的。 在 此外，对酵母信息素生物合成的了解可能使 酵母细胞在酵母细胞上生产具有医学价值的人类激素 商业规模。