MOLECULAR GENETICS OF THE DROSPHILA ZESTE LOCUS

果蝇 ZESTE 基因座的分子遗传学

• 批准号: 3280369
• 负责人: MICHAEL L GOLDBERG
• 金额: $ 16.6万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280369
• 关键词: DNA; DNA binding protein; Drosophilidae; alleles; chromosome aberrations; cytogenetics; endonuclease; gene expression; gene mutation; genetic manipulation; genetic models; genetic recombination; genetic transcription; immunofluorescence technique; immunoprecipitation; molecular cloning; molecular genetics; nucleic acid sequence; nucleic acid structure; protein structure function; radionuclides; transcription factor; yeasts

An investigation of the role of the Drosophila zeste gene product in synapsis-dependent gene expression (transvection) is proposed. The experiments described will extend previous findings that this protein binds to specific DNA sequences and acts as a transcription factor in vitro and cotransfection assays. The approach outlined first requires detailed study of the organization of functional domains of the zeste protein. This information will then be used in the design of model systems to test specific hypotheses of its activities in transvection. Using deletions of zeste sequences, domains of zeste required for its nuclear localization, possible multimerization or contacts with other proteins, cooperative DNA binding, and transcriptional activation properties will be identified. This work will be supplemented by an analysis of altered zeste proteins encoded by mutations with known genetic properties. Information generated in this manner should add to understanding of the molecular basis of dominance relationships between zeste alleles and the differing requirements for target gene dosage in the manifestation of their mutant phenotypes. Identification of regions of the zeste protein involved in particular aspects of its function will be exploited to create chimeric proteins useful for the establishment of a model system for transvection. Trials in both yeast and Drosophila will be performed to test detailed hypotheses for the role of the zeste gene product in this phenomenon. Possibilities to be investigated include: (a) zeste mediates the looping and pairing of DNA sequences; (b) zeste function requires synergistic interactions with other transcription factors, (c) zeste functions in a manner similar to other transcription factors, but unique elements of its structure are required for transvection, and (d) cooperative binding of zeste subunits is central to its function in transvection.

果蝇 zeste 基因产物在 提出了突触依赖性基因表达（横传）。 这 所描述的实验将扩展先前的发现，即该蛋白质结合 特定 DNA 序列并在体外充当转录因子 共转染测定。 首先概述的方法需要详细研究 zeste 蛋白功能域的组织。 这 然后信息将用于模型系统的设计以进行测试 其横传活性的具体假设。 使用 zeste 序列的删除，其所需的 zeste 结构域 核定位、可能的多聚化或与其他物质的接触 蛋白质、协同 DNA 结合和转录激活 属性将被识别。 这项工作将得到补充 分析由已知遗传突变编码的改变的zeste蛋白 特性。 以这种方式生成的信息应添加到 了解之间优势关系的分子基础 zeste等位基因和靶基因剂量的不同要求 其突变表型的表现。 鉴定特别涉及的zeste蛋白区域 其功能的各个方面将被用来创建嵌合蛋白 对于建立平移模型系统很有用。 试验于 酵母和果蝇都将被用来测试详细的假设 zeste 基因产物在这一现象中的作用。 的可能性 研究的内容包括：(a) zeste 介导 DNA 的循环和配对 序列； (b) zeste 功能需要与其他功能协同相互作用 转录因子，(c) zeste 的功能与其他因子类似 转录因子，但需要其结构的独特元件 对于横传，(d) zeste 亚基的协同结合是核心 其在平移中的功能。