Role of SR Proteins in Trans-splicing

超出提供的空间。这项提议的长期目标是确定SR蛋白在反式剪接中的作用。这将在从寄生线虫蛔虫衍生的实验系统中完成。我们的模型系统提供了在体外检测同一提取物中反式剪接和c/S剪接的独特能力。此外，提取物和剪接因子都可以从发育阶段的胚胎中制备出来。拟议的实验建立在我们在建立该系统在研究反式和c/S剪接之间的相互作用以及它在解决与SR蛋白和选择性剪接的发育调控相关的许多问题方面的有效性方面所取得的进展的基础上。我们还确定了反式剪接反应可以分为至少两个不同的阶段，从而使我们能够独立地检测SR蛋白在每个步骤中的参与情况。提出了三个具体的目标：1.研究反式剪接和顺式剪接的整合发育规律。2.确定U2 SnRNP加入底物前后SR蛋白参与反式剪接的条件。3.研究外显子定义在反式剪接中的作用及其与U1单链RNP的关系。彻底了解反式剪接反应可能有助于确定潜在的治疗干预点。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. The long term goal of this proposal is to determine the roles of SR proteins in trans-splicing. This will be accomplished in an experimental system derived from the parasitic nematode, Ascaris lumbricoides. Our model system offers the unique ability to assay both trans- and c/s-splicing in the same extracts in vitro. Additionally, both extracts and splicing factors can be prepared from developmentally staged embryos. The proposed experiments build upon the progress that we have made in establishing the utility of this system in examining the interplay between trans- and c/s-splicing as well as its applicability in addressing numerous questions related to the developmental regulation of both SR proteins and alternative splicing. We have also determined that the trans-splicing reaction can be separated into at least two distinct stages allowing us to assay the participation of SR proteins in each step independently. Three specific aims are proposed: 1. Examine the integrated developmental regulation of trans- and cis-splicing. 2. Determine the requirements for SR protein participation in trans-splicing before and after U2 snRNP addition to the substrate. 3. Investigate the role of exon definition and the involvement of U1 snRNP in trans-splicing. A thorough understanding of the trans-splicing reaction may lead to the identification of potential points of therapeutic intervention. PERFORMANCE SITE ========================================Section End===========================================