The molecular mechanism of pre-mRNA splicing

摘要 在所有真核生物中，前mRNA剪接是基因表达所必需的，剪接错误导致基因 疾病和许多其他疾病。深入了解前信使核糖核酸的分子机制 剪接有可能为人类疾病治疗提供有用的方法。内含子的剪接是 通过剪接体催化的两个酯交换反应进行，剪接体是一种大的RNA/蛋白质 由5个SnRNP(U1、U2、U4、U5、U6)和许多非SnRNP相关蛋白因子组成的复合体。这个 剪接体在剪接循环中经历戏剧性的变化，产生E，A，前-B，B，Bact，B*，C，C*，P， 和ILS复合体。在过去的40年里，对遗传、生化和结构的研究产生了 关于剪接机制的大量信息，但仍然存在重大的知识差距。我们的目标是 未来5年是使用结构、生化和遗传/基因组学方法的组合来 了解备选5‘识别的机制和调控，外显子定义的机制， 转录和剪接偶联的机制和功能。我们预计这些项目将填补 几个重要的知识空白，并促进了我们对剪接机制的理解。

Summary Pre-mRNA splicing is essential for gene expression in all eukaryotes and errors in splicing cause genetic disorders and many other diseases. A thorough understanding of the molecular mechanisms of pre-mRNA splicing has the potential to provide useful approaches for human disease therapy. The splicing of introns is carried out through two transesterification reactions catalyzed by the spliceosome, a large RNA/protein complex composed of five snRNPs (U1, U2, U4, U5, U6) and many non-snRNP related protein factors. The spliceosome undergoes dramatic changes in a splicing cycle, generating the E, A, Pre-B, B, Bact, B*, C, C*, P, and ILS complexes. Genetic, biochemical, and structural studies in the past four decades have generated tremendous information on the mechanism of splicing, but significant knowledge gaps remain. Our goal in the next 5 years is to use a combination of structural, biochemical, and genetic/genomics approaches to understand the mechanism and modulation of alternative 5’ ss recognition, the mechanism of exon definition, and the mechanism and function of transcription and splicing coupling. We envision that these projects will fill several significant knowledge gaps and advance our understanding of the mechanism of splicing.