GENETIC ANALYSIS OF YEAST GAL GENE REGULATION

作用于酵母的GAL基因的调节回路 S.酿酒酵母在遗传上是明确的。 核心利益是 GAL 4编码的调节蛋白 激活GAL基因的转录。 这种蛋白质与 位于被调节的每个基因上游的序列; DNA结合后发生的事件，导致 转录的激活是未知的。 也知之甚少 是负责GAL的分解代谢物抑制的因素 由葡萄糖生长引起的基因。 的长期目标 一项拟议的研究是了解蛋白质如何与每个 其他的和GAL控制区调节转录。 我们将集中在三种类型的蛋白质：1）GAL 4编码的 蛋白质，2）与GAL 4蛋白质相互作用的蛋白质，和3） 负责GAL基因的分解代谢物抑制的蛋白质。 具体目标是： 1)为了定位GAL 4编码蛋白的功能区域， 通过鉴定影响DNA结合的突变， 转录激活和该蛋白的其他功能。 因为在这个大的区域中功能区域的位置 蛋白质定义不清，那么体内的几种蛋白质将被 在更精确定义的区域进行。 2)鉴定与GAL 4蛋白相互作用的蛋白质。 GAL 4蛋白可能通过以下方式激活转录： 与转录附近的蛋白质相互作用 起始位点，但编码这些蛋白质的基因没有 是转录起始位点，但编码这种基因的基因 蛋白质尚未被鉴定。 基因选择 这里提出的目的是鉴定这些基因。 3)为了阐明分解代谢物阻遏的机制， 通过分离和表征突变体的GAL基因 在使用选择方案的这种调节电路中有缺陷 专门为此开发的。 4)开发一种基因分析方法， 结合蛋白 这些方法将被应用到本研究中 GAL 4蛋白，以及其他DNA结合蛋白。

The regulatory circuit that acts upon the GAL genes of the yeast S. cerevisiae is well defined genetically. Of central interest is the mechanism by which the GAL4-encoded regulatory protein activates transcription of the GAL genes. This protein binds to sequences located upstream of each gene that is regulates; the events that occur subsequent to DNA binding that lead to activation of transcription are unknown. Also poorly understood are the factors responsible for catabolite repression of the GAL genes caused by growth on glucose. The long-term goal of the proposed research is to learn how proteins interact with each other and with the GAL control regions to regulate transcription. We will focus on three types of proteins: 1) the GAL4-encoded protein, 2) proteins that interact with the GAL4 protein, and 3) proteins responsible for catabolite repression of the GAL genes. The specific aims are: 1) To locate functional regions of the GAL4-encoded protein by identifying mutations that affect the DNA binding, transcription activation, and other functions of this protein. Because the location of functional regions in this large protein are poorly defined, several in vivo will then be carried out on more precisely defined regions. 2) To identify proteins that interact with the GAL4 protein. The GAL4 protein likely activates transcription by interacting with proteins that act close to the transcription initiation site, but genes encoding such proteins have not been transcription initiation site, but genes encoding such proteins have not been identified. The genetic selections proposed here are aimed at identification of these genes. 3) To elucidate the mechanism of catabolite repression of the GAL genes by isolating and characterizing mutants defective in this regulatory circuit, using selection schemes specifically developed for this purpose. 4) To develop a genetic method for the analysis of DNA binding proteins. These methods will be applied to the study of GAL4 protein, as well as other DNA binding proteins.