YEAST PHEROMONE GENES STRUCTURE, FUNCTION, AND CONTROL

α因子是一种由十三个氨基酸组成的寡肽信息素， 由酵母的一种单倍体细胞类型（MAT α细胞）产生。 相反交配类型的细胞（MAT α）通过以下方式对α因子作出反应： 在细胞周期的G1期停滞，在该点细胞是 能够交配。 克隆的α-因子结构基因（MAF）的分析 表明该基因编码前聚α因子 前体与四个串联重复的α因子，这需要几个 用于生产成熟α-因子的加工步骤。 杂交 利用MAF α DNA的研究表明，可能存在额外的 酵母基因组中的α因子结构基因（MAF α '）。 目标 这个建议的目的是回答以下问题：1）α因子是 交配所需的？ 2)α因子前体本身是否发挥了作用 在交配中 3)有多少活跃的α因子结构基因存在于 酵母基因组 4)如果不止一个人在场，他们都需要吗？ 为了交配 5)什么步骤涉及在加工和分泌 α-因子和前体的哪些区域是这些所需的 步骤？ 6)α因子生产的细胞类型特异性如何 决心？ 将使用的技术包括克隆和DNA 对MAF α基因进行测序以确定其结构， 体内活性试验，体外诱变，然后基因 替换，以评估表型（即，确定 α因子及其前体在交配中的作用不同 加工和分泌途径中的前体部分），以及 将lacZ融合物构建到这些基因的5 ′侧翼区， 随后进行缺失分析，以确定基因的区域 参与调控和表达。 此外，还将尝试 克隆α-因子结构基因。 α因子类似于 MAT Alpha细胞产生的信息素。 它是一种十一种氨基酸 寡肽，其以两种形式存在，仅相差一个氨基酸 替代。 我的研究结果表明MAF α基因编码一种 多α因子前体表明，两种形式的α因子可能 在编码多α因子前体的单个基因中编码。 将对α因子结构基因进行测序，以确定是否 情况就是这样，类似于阿尔法因素的问题将 接受检查。

Alpha-factor is an oligopeptide pheromone of thirteen amino acids which is produced by one of the haploid cell types of yeast (MAT Alpha cells). Cells of the opposite mating type (MAT Alpha) respond to Alpha-factor by arresting in the G1 phase of the cell cycle, the point at which cells are able to mate. Analysis of a cloned Alpha-factor structural gene (MAF Alpha) indicates that this gene encodes a pre-pro-poly-Alpha-factor precursor with four tandem repeats of Alpha-factor, which requires several processing steps for the production of mature Alpha-factor. Hybridization studies utilizing the MAF Alpha DNA indicate that there may be additional Alpha-factor structural genes (MAF Alpha') in the yeast genome. The aims of this proposal are to answer the following questions: 1) Is Alpha-factor required for mating? 2) Does the Alpha-factor precursor itself play a role in mating? 3) How many active Alpha-factor structural genes are present in the yeast genome? 4) If more than one is present, are they all required for mating? 5) What steps are involved in processing and secretion of Alpha-factor and which regions of the precursor are required for these steps? 6) How is the cell-type-specificity of Alpha-factor production determined? The techniques to be used include the cloning and DNA sequencing of the MAF Alpha' genes in order to determine their structure, in vivo tests of their activity, in vitro mutagenesis followed by gene replacement in order to assess phenotypes (i.e. to determine the roles of Alpha-factor and its precursor in mating and the role of the different parts of the precursor in the processing and secretion pathway), and the construction of lacZ fusions to the 5' flanking regions of these genes, followed by deletion analysis, to determine the regions of the gene involved in regulation and expression. Also, an attempt will be made to clone the Alpha-factor structural gene. Alpha-factor is the analogous pheromone produced by MAT Alpha cells. It is an eleven amino acids oligopeptide, which exists in two forms which differ by a single amino acid substitution. My results indicating that the MAF Alpha gene encodes a poly-Alpha-factor precursor suggest that the two forms of Alpha-factor may be encoded in a single gene which codes for a poly-Alpha-factor precursor. The Alpha-factor structural gene will be sequenced to determine whether this is the case and questions similar to those posed for Alpha-factor will be examined.