MOLECULAR BASIS OF GENE EXPRESSION IN YEAST

酵母基因表达的分子基础

• 批准号: 3295207
• 负责人: Elizabeth Joan Grayhack
• 金额: $ 14.1万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1990-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3295207
• 关键词: DNA; Saccharomyces; cell type; fungal genetics; gene expression; genetic transcription

In the yeast S. cerevisiae, the alpha 1 protein is responsible for expression of a set of cell-type specific genes, the alpha-specific genes. Three such genes, MF(alpha)1, and MF(alpha)2, and STE3, have been identified and code for proteins required for the specialized functions of the alpha cell type. Our goal is to understand the molecular basis for two aspect of the alpha 1 protein's regulatory activity: 1) the specificity of alpha 1 protein for the alpha-specific genes and 2) the mechanism by which alpha 1 protein implements its activation of gene expression. We will use the results of our analysis of the action of the alpha 1 protein on the MF(alpha)1, promoter to understand how the separate elements of a yeast promoter are coordinated. To analyze the activity of the alpha 1 protein, we will begin by purifying the alpha 1 protein. We will test the idea that the alpha 1 protein is specific for activating expression of the MF(alpha)1 gene because it binds to sequences near this gene. If so, is it solely a property of alpha 1 protein or are other yeast proteins involved? Since the promoter is a multi-component system, it seems likely that alpha 1 protein exerts its effects by interacting with other transcription components; by identifying and characterizing these interacting proteins we may learn whether or not proteins that bind to separate promoter elements interact with each other. To understand how the separate sequence elements (and their associated proteins) are coordinated in promoter function, we will identify other proteins that interact with the MF(alpha)1 promoter. We will drive formation of a transcription complex with the DNA binding proteins and alpha 1 protein and will attempt to reproduce specific alpha 1 dependent transcription in vitro. In addition to our interest in how the alpha 1 protein acts, we are also interested in how the alpha 1 protein is regulated. Some strains of yeast can switch mating type frequently; the rapid acquisition of specialized cell function by the newly switched cell is suggestive evidence that the old cell type regulators, i.e., alpha 1 (and others) are rapidly inactivated after the cell type switch occurs. We propose to examine the inactivation of the alpha 1 protein to discern how this putative activity is regulated.

在酵母S.α 1蛋白负责 一组细胞类型特异性基因的表达，α-特异性 基因. 三个这样的基因，MF（alpha）1，MF（alpha）2，和STE 3， 已经被鉴定并编码了 阿尔法细胞的特殊功能 我们的目标是 了解阿尔法1的两个方面的分子基础 蛋白的调节活性：1）α 1蛋白的特异性 2）α-特异性基因的机制， 1蛋白实现其对基因表达的激活。 我们将 利用我们对α 1蛋白作用的分析结果， 对MF（alpha）1启动子的研究，以了解如何分离 酵母启动子的元件是配位的。 为了分析α 1蛋白的活性，我们将开始 纯化α 1蛋白。 我们将验证阿尔法狼 1蛋白特异性激活MF（alpha）1的表达 因为它与这个基因附近的序列结合。 如果是的话， 仅仅是α 1蛋白的性质，或者是其它酵母蛋白 牵涉其中？ 由于启动子是多组分系统， α 1蛋白似乎是通过与 与其他转录成分;通过识别和 描述这些相互作用的蛋白质，我们可以了解， 而不是与单独的启动子元件结合的蛋白质 彼此之间 要了解单独的序列元素（及其 相关蛋白）在启动子功能中协调，我们将 鉴定与MF（alpha）1相互作用的其他蛋白质 启动子 我们将驱动转录复合体的形成 DNA结合蛋白和α 1蛋白， 试图复制特定的α 1依赖性转录， 体外 除了我们对α 1蛋白如何起作用的兴趣外， 也对α 1蛋白是如何调节的感兴趣。 一些 酵母菌株可以频繁地转换交配型， 通过新切换的细胞获得专门的细胞功能 提示性证据表明，旧的细胞类型调节器，即，阿尔法 1（和其他）在细胞类型转换后迅速失活 发生。 我们建议检查α 1的失活， 蛋白质来辨别这种假定的活性是如何调节的。