YEAST PEPTIDE HORMONE SYNTHESIS AND SIGNAL TRANSDUCTION

酵母肽激素的合成和信号转导

• 批准号: 2173812
• 负责人: Jeremy W. Thorner
• 金额: $ 41.07万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1975
• 资助国家: 美国
• 起止时间: 1975-01-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2173812
• 关键词: DNA replication; Saccharomyces; Saccharomyces cerevisiae; adenylate cyclase; affinity chromatography; binding proteins; biological signal transduction; carbon; cell cell interaction; cell cycle; cell membrane; eukaryote; fungal genetics; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; glycoproteins; high performance liquid chromatography; hormone regulation /control mechanism; iodine; membrane activity; messenger RNA; molecular cloning; oligopeptides; peptide hormone; peptide hormone biosynthesis; pheromone; phosphorylation; radioimmunoassay; radionuclides; receptor; recombinant DNA; structural genes; temperature sensitive mutant

The objective of this research is to learn the precise molecular details of both the biosynthesis of peptide hormones and the mechanism of action of peptide hormones. Secreted bioactive peptides (peptide hormones, growth factors, neuroregulatory pep- tides) are indispensable agents of the intercellular interactions that control both embryonic development and the coordination and integration of life functions in the adult. Numerous human diseases are associated with the under- (or over-) production of regulatory peptides, or with the inability to respond properly to such signals. Even the unicellular eukaryotic microorganism Saccharomyces cerevisiae (baker's yeast) secretes and responds to peptide hormones ("mating pheromones")- Because incisive genetic and biochemical studies can be performed in yeast cells, particularly those that utilize recombinant DNA, study of peptide hormone biogenesis and signal transduction in yeast may provide useful insights for understanding the molecular basis of diseases that affect the human neuroendocrine system. Four specific areas of investigation are proposed: (1) The molecular basis of cell type-specific gene expression will be studied by examining the transcriptional regulation of the MF a1 gene (which encodes a precursor, prepro-a-factor, of one of the mating pheromones)- Genetic and biochemical experiments are proposed for further defining both the cis-acting sites and the trans-acting factors responsible for the control of this gene, and for exploring the role of a particular "zinc finger"- containing transcription factor (STE5 gene product). (2) The enzymic basis of pre= cursor maturation at sites comprised of pairs of basic residues will be explored by determination of the three-dimensional structure of a yeast precursor cleaving enzyme (KEX2 gene product) with this specificity and by using probes based on this yeast gene to isolate its mammalian homolog. (3) How a plasma membrane receptor acts as a ligand-triggered molecular switch and how its function is regulated during desensitization will be elucidated using the cloned alpha-factor receptor gene (STE2), its coupled G a subunit (GPAl gene product), and a regulatory factor (SST2 gene product) of as yet unknown function. (4) Hormonal induction of gene transcription will be studied by genetic and biochemical experiments designed to identify and characterize the protein factors that interact with an upstream pheromone-response element (PRE).

这项研究的目的是学习精确的分子 肽激素和肽激素的生物合成的细节和 肽激素的作用机理。 分泌的生物活性 肽（肽激素，生长因子，神经调节性PEP- 潮汐）是细胞间相互作用的必不可少的药物 控制胚胎发展和协调和 成人生活功能的整合。 许多人 疾病与未经（或过度）的生产有关 监管肽，或者无法正确做出反应 这样的信号。 甚至单细胞真核生物微生物 酿酒酵母（贝克的酵母）分泌并回应 肽激素（“交配信息素”） - 因为尖锐的遗传 并且可以在酵母细胞中进行生化研究， 特别是那些利用重组DNA的人，研究肽 酵母中的激素生物发生和信号转导可能会提供 理解疾病分子基础的有用见解 这会影响人类神经内分泌系统。 提出了四个特定的调查领域：（1） 细胞类型特异性基因表达的分子基础将是 通过检查MF A1的转录调控来研究 基因（编码一个前体的前体，即prepro-a因子，其中之一 交配信息素） - 遗传和生化实验是 提议进一步定义顺式作用位点和 负责控制该基因的跨作用因子， 并探索特定的“锌指”的作用 - 包含转录因子（Ste5基因产物）。 （2） PRE =光标成熟的酶基础在由由 通过确定，将探索成对的基本残留物 酵母前体切割酶的三维结构 （Kex2基因产品）具有这种特异性并使用探针 基于该酵母基因，以分离其哺乳动物同源物。 （3） 质膜受体如何充当配体触发 分子开关及其功能如何调节 使用克隆的α因子阐明脱敏 受体基因（Ste2），其耦合G A亚基（GPAL基因 产品）和一个调节因素（SST2基因产物） 未知功能。 （4）基因转录的激素诱导 将通过设计的遗传和生化实验来研究 识别和表征相互作用的蛋白质因子 带有上游信息素反应元件（PRE）。