Alternative pre-mRNA Splicing in Drosophila

描述(由申请人提供)： 项目摘要/摘要果蝇的Pre-mRNAs剪接。在后生动物中，D.Rio-PI选择性的Pre-mRNA剪接是调节基因表达的重要机制。的确，RNA处理是基因组信息传递到蛋白质组信息的管道。由于大多数真核基因是分裂的，具有选择性剪接的潜力，细胞、组织和生物之间蛋白质组多样性的急剧增加是选择性剪接的直接结果。事实上，现在已经认识到，已知的人类和小鼠疾病基因突变中有30%-40%会影响剪接过程。因此，了解内含子是如何识别的，以及选择性剪接模式是如何建立的，可能会允许进行治疗干预。剪接沉默是在后生动物中产生组织或细胞类型特异性剪接模式的主要类型的RNA控制元件。我们之前的工作主要集中在组织特异性的果蝇P元件剪接沉默元件的表征上。我们小组最近的工作集中在选择性剪接的组织特异性调控以及hnRNP蛋白、PSI和hnRNP A/B家族蛋白hrp48、40、38和36如何导致剪接沉默功能。我们希望将这些研究扩展到研究U2AF的果蝇副基因LS2的功能，该基因具有剪接抑制因子的功能。在之前R21资助的基础上，我们将继续研究microRNA途径蛋白AGO-2在替代剪接途径中的作用。最后，使用各种方法，我们想要识别和功能表征由多种剪接因子控制的新型细胞剪接消声器。这一提议的一个优势是基因组规模小，注释良好的果蝇基因组，允许全面分析备选剪接模式和RNA结合蛋白在全基因组的分布。由于大多数真核基因是分裂的，具有选择性剪接的潜力，细胞、组织和生物之间蛋白质组多样性的增加是选择性剪接的直接结果。由于参与控制选择性剪接模式的蛋白质因子在人类中具有同源性，而且果蝇和人类一样具有广泛的选择性剪接，因此该系统所获得的结果将直接与人类健康相关并应用于此。 公共卫生相关性： 果蝇D rio-Pi选择性前mRNA剪接是调节后生动物基因表达的一种常见机制。由于大多数真核基因是分裂的，具有选择性剪接的潜力，细胞、组织和生物之间蛋白质组多样性的急剧增加是选择性剪接的直接结果。事实上，现在已经认识到，已知的人类和小鼠疾病基因突变中有30%-40%影响剪接过程，由于控制选择性剪接模式的蛋白质因子在人类中具有同源性，而且果蝇与人类一样具有广泛的选择性剪接，因此在该系统中获得的结果将直接与人类健康相关并应用于人类健康。

DESCRIPTION (provided by applicant): PROJECT SUMMARY / ABSTRACT Alternative pre-mRNA splicing in Drosophila. D. Rio - PI Alternative pre-mRNA splicing is an important mechanism for regulating gene expression in metazoans. Indeed, RNA processing is a conduit through which genomic information is transferred to proteomic information. Because most eukaryotic genes are split and have the potential for alternative splicing, a dramatic increase in proteomic diversity among cells, tissues and organisms is a direct consequence of alternative splicing. In fact, it is now recognized that 30-40% of the known human and mouse disease gene mutations affect the splicing process. Thus, understanding how introns are recognized and how patterns of alternative splicing are set up may allow therapeutic intervention. Splicing silencers are a major type of RNA control element generating tissue- or cell type-specific splicing patterns in metazoans. Our previous work has focused on characterization of the tissue-specific Drosophila P element splicing silencer element. Recent work from our group has focused on tissue-specific regulation of alternative splicing and how the action of hnRNP proteins, PSI and the hnRNP A/B family proteins, hrp48, 40, 38 and 36 results in splicing silencer function. We wish to extend these studies to investigate the function of a Drosophila paralog of U2AF, called LS2, that functions as a splicing repressor. Building on previous R21 funding, we will continue investigations of the microRNA pathway protein, Ago-2, in alternative splicing pathways. Finally, using a variety of approaches, we want to identify and functionally characterize new cellular splicing silencers that are controlled by multiple splicing factors. One advantage this proposal has is the small genome size and well-annotated Drosophila genome that allows a comprehensive analysis of both alternative splicing patterns and the genome-wide distribution of RNA binding proteins. Because most eukaryotic genes are split and have the potential for alternative splicing, increased proteomic diversity among cells, tissues and organisms is a direct consequence of alternative splicing. Because the protein factors involved in controlling patterns of alternative splicing have homologs in humans and like humans Drosophila has extensive alternative splicing, results obtained in this system will have direct relevance and application to human health. PUBLIC HEALTH RELEVANCE: PROJECT NARRATIVE Alternative pre-mRNA splicing in Drosophila D Rio - PI Alternative pre-mRNA splicing is a common mechanism for regulating gene expression in metazoans. Because most eukaryotic genes are split and have the potential for alternative splicing, a dramatic increase in proteomic diversity among cells, tissues and organisms is a direct consequence of alternative splicing. In fact, it is now recognized that 30-40% of the known human and mouse disease gene mutations affect the splicing process and because the protein factors involved in controlling patterns of alternative splicing have homologs in humans and like humans Drosophila has extensive alternative splicing, results obtained in this system will have direct relevance and application to human health.