Regulation of Sex-lethal pre-mRNA splicing during Drosophila development

果蝇发育过程中性致死性前体 mRNA 剪接的调控

基本信息

批准号：
7321870
负责人：
HELEN Karen SALZ
金额：
$ 30.9万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-05-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7321870
关键词：
3&apos Splice Site Alternative Splicing Amino Acid Motifs Animals Biochemical Biochemical Genetics Biochemistry Biological Models Compatible Complex Couples Data Development Disease Drosophila genus ELAV protein Embryo Equilibrium Event Exons Female Gene Expression Genes Genetic Genetic Screening Genetic Transcription Germ Cells Goals Hela Cells Heterogeneous Nuclear RNA Histocompatibility Testing Human Human Biology In Vitro Malignant Neoplasms Mediating Modeling Molecular Mutation Nuclear Extract Numbers Pattern Play Process Protein Binding Protein Family Protein Splicing Proteins RNA RNA Processing RNA Splicing RNA-Binding Proteins Recruitment Activity Regulation Repression Research Personnel Role Site Specificity Spliceosome Assembly Pathway Spliceosomes Staging Switch Genes System Testing Time Tissues Transcriptional Regulation Translations U1 Small Nuclear Ribonucleoprotein Work base fly genetic regulatory protein human disease in vivo insight mRNA Precursor male member neuronal cell body novel programs protein function research study sex tissue/cell culture

项目摘要

DESCRIPTION (provided by applicant): Tissue specific alternative splicing plays a prominent role in regulating gene expression in metazoans. Although the molecular events required to splice a simple pre-mRNA in vitro are fairly well understood, we remain largely ignorant of how the complex process of splicing is regulated in vivo. An understanding of how this process works is of fundamental importance since errors in RNA splicing are associated with a variety of human diseases, including cancer. The long-term goal of this project is to understand regulation by exon skipping, one of the most common forms of alternative splicing in humans. Specifically, we study the splicing regulation of the Drosophila developmental switch gene Sex-lethal (Sxl). Based on our earlier studies, we propose a novel mechanism by which the female specific product of the Sxl gene controls its own splicing pattern by interacting with, and inactivating, core components of the spliceosome. As outlined in the proposal, we are continuing to exploit Drosophila genetics to identify and delineate the function of proteins that participate in this biologically important example of splicing regulation. We are also using biochemical approaches to test the models that are known to be compatible with what we observe in vivo. Lastly, we are investigating, for the first time, whether the mechanism utilized to control Sxl is tissue-, as well as sex-specific. The combined data from these studies will provide insights into the complexities associated with splicing regulation that simply cannot be obtained via studies carried out solely in tissue culture cells. Due to the remarkable conservation of the splicing machinery from humans to flies, the paradigms we develop using the very tractable Drosophila system are relevant to human biology.

描述（由申请人提供）：组织特异性选择性剪接在调节后生动物的基因表达中发挥着重要作用。尽管在体外剪接简单的前mRNA所需的分子事件已被很好地理解，但我们仍然对体内如何调节复杂的剪接过程一无所知。了解这一过程的工作原理至关重要，因为 RNA 剪接错误与多种人类疾病（包括癌症）相关。该项目的长期目标是了解外显子跳跃的调节，外显子跳跃是人类最常见的选择性剪接形式之一。具体来说，我们研究了果蝇发育开关基因性致死（Sxl）的剪接调控。基于我们早期的研究，我们提出了一种新的机制，通过该机制，Sxl 基因的女性特异性产物通过与剪接体的核心成分相互作用并使其失活来控制其自身的剪接模式。正如提案中所述，我们正在继续利用果蝇遗传学来识别和描述参与这一重要的剪接调节示例的蛋白质的功能。我们还使用生化方法来测试已知与我们在体内观察到的情况兼容的模型。最后，我们首次研究用于控制 Sxl 的机制是否具有组织特异性和性别特异性。这些研究的综合数据将提供对与剪接调节相关的复杂性的见解，而这些复杂性是无法通过仅在组织培养细胞中进行的研究获得的。由于从人类到果蝇的剪接机制具有显着的保守性，我们使用非常容易处理的果蝇系统开发的范例与人类生物学相关。