Genetic Analysis of Alpha-Tubulin Function in Drosophila

果蝇α-微管蛋白功能的遗传分析

• 批准号: 9004477
• 负责人: Kathleen Matthews
• 金额: $ 20万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-15 至 1995-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9004477&HistoricalAwards=false
• 关键词: Genetic; Analysis; Alpha; Tubulin; Function

Tubulin plays a crucial role in morphogenesis as the primary component of microtubules. We propose to address two basic questions about these important structural proteins using the four a-tubulins of Drosophila: are any of the a-tubulin isotypes functionally specialized, and what regulatory mechanisms are responsible for achieving the correct ratios and concentrations of a-tubulins isotypes in each cell type? The multifunctionality of each a-tubulin isotype will be tested by determining the ability of one isotype to rescue developmental defects associated with mutations in another isotype. Transgenic strains carrying ectopic copies of each a-tubulin gene and mutations in two of the a- tubulins are available. An attempt will be made to generate mutations in the two remaining genes using P-element mutagenesis and PCR, an approach that allows the recovery of mutations with no detectable phenotype. Regulatory mechanisms operating to maintain tissue-appropriate a-tubulin pools will be identified by examining the ability of cells to 1) eliminate misexpressed a-tubulin isotypes produced from chimeric genes, and 2) respond to overexpression of appropriate isotypes from supernumerary tubulin genes. Genes encoding transacting a-tubulin regulatory factors will be sought by reverting the female sterility associated with overexpression of a maternal tubulin. The protein tubulin is an important structural component of most types of cells. It exists in a number of different forms, some present in all cells, others present only in certain specialized types of cells. The different forms of tubulin presumably serve different functions, but very little is known about what these functions are. This is a proposal to investigate the different functions of the different forms of tubulin.

微管蛋白在形态发生中起着重要作用， 微管的组成部分。 我们建议解决两个基本问题 关于这些重要的结构蛋白质的问题， 果蝇的α-微管蛋白：是任何α-微管蛋白同种型 功能专业化，以及什么样的监管机制 负责实现正确的比例和浓度， 每种细胞类型中的α-微管蛋白同种型？ 的多功能性 每种α-微管蛋白同种型将通过测定 一种同种型，以挽救与 另一种同种型的突变。 转基因菌株携带异位 每个α-微管蛋白基因的拷贝和两个α-微管蛋白基因的突变， 微管蛋白是可用的。 将尝试生成 使用P元件诱变在剩余的两个基因中突变 和PCR，一种允许恢复突变的方法， 可检测的表型 监管机制的运作， 通过检测细胞中的细胞因子， 细胞1）消除错误表达的α-微管蛋白 由嵌合基因产生的同种型，和2）响应于 多余微管蛋白适当同种型的过表达 基因. 编码反式α-微管蛋白调节因子的基因 将通过恢复与之相关的女性不育来寻求 母体微管蛋白的过度表达。 蛋白质微管蛋白是大多数细胞的重要结构组分， 细胞类型。 它以多种形式存在，有些 存在于所有细胞中，其他的只存在于某些特定的细胞中。 细胞类型。 不同形式的微管蛋白可能 不同的功能，但人们对这些功能知之甚少 功能是。 这是一个调查不同 不同形式微管蛋白的功能。