YEAST PHEROMONE GENES STRUCTURE, FUNCTION, AND CONTROL

α-因子是一种由13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(α细胞)产生的。 相反交配型的细胞(MAT Alpha)通过以下方式对Alpha因子做出反应 停滞在细胞周期的G1期，细胞处于 能够交配。一个克隆的α-因子结构基因(MAF)的分析 α)表明该基因编码一种前多聚α因子 具有四个α-因子串联重复序列的前体，这需要几个 成熟α-因子的生产工艺步骤。杂交 利用MAF阿尔法DNA的研究表明，可能还有其他 酵母基因组中的α因子结构基因(MAFα‘)。目标 这一建议是回答以下问题：1)是阿尔法因子 交配所需？2)阿尔法因子前体本身起作用吗？ 在交配中？3)有多少活跃的阿尔法因子结构基因存在于 酵母基因组？4)如果存在不止一个，它们都是必需的吗 对于交配？5)什么步骤涉及加工和分泌 α因子以及这些需要前体的哪些区域 步骤？6)阿尔法因子产生的细胞类型特异性如何 有决心吗？将使用的技术包括克隆和DNA 对MAF Alpha‘基因进行测序以确定其结构， 它们的体内活性测试、体外诱变和基因突变 替换，以评估表型(即确定 α-因子及其前体在交配中的作用 前体在加工和分泌途径中的部分)，以及 构建与这些基因5‘侧翼区的LacZ融合， 然后进行缺失分析，以确定基因的区域 参与调控和表达的。此外，还将尝试 克隆α因子结构基因。α因子是类似的 垫状阿尔法细胞产生的信息素。它由十一种氨基酸组成 寡肽，以两种形式存在，只有一种氨基酸不同 换人。我的结果表明MAF Alpha基因编码一个 多α因子前体提示两种形式的α因子可能 被编码在单个基因中，该基因编码多阿尔法因子前体。 将对阿尔法因子结构基因进行测序，以确定 这种情况和问题类似于对阿尔法因子Will提出的问题 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.