REGULATION OF SEX-SPECIFIC RNA SPLICING IN DROSOPHILA

果蝇性别特异性 RNA 剪接的调控

• 批准号: 3305296
• 负责人: RODNEY N NAGOSHI
• 金额: $ 13.5万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3305296
• 关键词: Drosophilidae; RNA binding protein; RNA splicing; crosslink; developmental genetics; genetic mapping; genetic regulation; genetic regulatory element; messenger RNA; molecular cloning; mutant; protein sequence; sex determination; transcription factor

This proposal addresses the general question of how developmental processes are regulated at the level of gene expression. The specific goal is to investigate the mechanisms by which alternative RNA processing controls the sex-specific expression of a gene that regulates somatic sexual differentiation in Drosophila melanogaster. The doublesex (dsx) gene acts as a "genetic switch"; it is expressed in either a male- or female-specific mode, the choice of which directs the course of somatic sexual development. The regulation of dsx occurs through sex-specific RNA processing. To understand how this regulation occurs, a three-stage strategy is proposed: (i) Identify and characterize the dsx sequences required in cis for sex- specific RNA splicing. This will be done by in vivo studies using dsx minigene constructs. The effect of site-directed mutations on the minigene splicing pattern will define the organization and behavior of these sites. (ii) Determine how these sites function, i.e., do they bind trans-acting regulatory factors and are they involved in important secondary structures? These studies will utilize RNA-protein binding assays including gel mobility shift and protection experiments. (iii) Isolate and characterize the trans-acting factors. The proposed research will contribute to our understanding of a developmental process that is fundamental to all sexually reproducing organisms. Furthermore, the insights gained on the regulation of RNA processing will have substantial relevance to the study of a number of genes that are regulated in a similar manner. These include genes of major medical importance involving such processes as oncogenesis, viral regulation, and the immune system, among others.

该提案解决了发展过程的一般问题 在基因表达水平上受调节。 具体目标是 研究替代RNA处理控制的机制 调节躯体性爱的基因的性别表达 果蝇的分化Melanogaster。 Doublesex（DSX）基因作用 作为“遗传转换”；它以男性或女性特异性表示 模式，指导躯体性发展过程的选择。 DSX的调控是通过性别特异性的RNA处理发生的。 到 了解该法规是如何进行的，提出了三阶段的策略： （i）识别和表征CI中所需的DSX序列的性别 - 特定的RNA剪接。 这将通过使用DSX的体内研究来完成 微型结构。 位置突变对微基因的影响 剪接模式将定义这些站点的组织和行为。 （ii）确定这些站点的功能，即它们是否结合反式作用 监管因素，它们是否参与重要的二级结构？ 这些研究将利用包括凝胶在内的RNA-蛋白结合测定 移动性转移和保护实验。 （iii）孤立和特征 跨作用因素。 拟议的研究将有助于我们对 发展过程对于所有性繁殖至关重要 有机体。 此外，对RNA的调节的见解 处理将与许多研究具有很大的相关性 以类似方式调节的基因。 这些包括主要的基因 医学重要性涉及诸如肿瘤发生，病毒的过程 监管和免疫系统等。