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: 3270737
• 负责人: Jeremy W. Thorner
• 金额: $ 16.13万
• 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/3270737
• 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.

酵母的两种单倍体细胞类型（a和α） 酿酒酵母各自分泌特异性的肽酶样分子， 触发事件导致细胞的融合（“交配”），相对而言， 简单的发育顺序。 因为深刻的遗传学和生物化学 研究可以在酵母细胞中进行，特别是那些利用 重组DNA，这可能是有可能了解精确的分子细节， 肽激素的生物合成和模式的行动，通过检查酵母 交配信息素（a因子和α因子）。 拟采取的办法 包括：（1）在体外改变克隆的 α-因子和α-因子基因以及与指示剂的新基因融合体 酶，以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， 与克隆的基因结合，以提供探针和底物 分析合成、膜转位、翻译后 信息素前体蛋白的加工和分泌运输 （前原-α-因子和前原-α-因子）;（3）[125 I]-标记的信息素， 反应性信息素衍生物的共价交联，亲和性 使用基质结合信息素的色谱法， α-因子受体（STE 2）和腺苷酸环化酶（CDC 35），以及克隆的 STE 5基因，以鉴定信息素受体，并研究其作用机制。 环AMP合成的信息素调节;和，（4）转录作图， 基因破坏和DNA测序，以表征结构和 克隆基因的功能，其表达受信息素控制。 在多细胞生物中，肽类激素负责细胞间的相互作用。 在胚胎发育期间和发育过程中所需的通信 成年人生活功能的协调。 生产不足或生产过剩 特别是肽激素，或无法感觉到它们的存在， 与许多人类疾病相关（例如侏儒症、库欣氏病 疾病和糖尿病）。 对《公约》基本方面的理解 肽激素的合成和释放，以及这些激素的调节 过程，以及其作用机制的生化基础， 是有效诊断和治疗此类疾病所必需的。 在 此外，酵母信息素的生物合成的知识可以允许 通过酵母细胞在一种培养基上生产具有医学价值的人类激素 商业规模。