Structural Study on the early stage spliceosomal complex

真核生物转录后事件，如剪接、转运和mrna的生命周期调节，对基因表达和细胞分化的调节是重要的。pre-mRNA的剪接是一个重要的事件，它在RNA被翻译成蛋白质之前去除非编码RNA的片段。在剪接反应的早期阶段，5‘剪接位点必须被U1 snRNP识别，3’剪接位点必须被U2AF蛋白识别，分支位点必须被U2 snRNP识别。在这项研究中，我们研究了参与这些成分的蛋白质的结构。1.SURP结构域主要存在于剪接机制中（如U2 snRNP）。我们测定了SF4蛋白中SURP结构域的结构。2.我们试图重建p14-SAP155复合体。通过核磁共振法证实了该配合物的成功重构。3.此外，对于U2AF蛋白的结构研究，我们可以建立u2af4的重组方案。U1C是U1 snRNP的一个组成部分。为了获得U1-C蛋白相互作用的信息，我们研究了U1-C蛋白n端（1-65）片段的结构。用同核磁共振法测定了U1-C（1-61）蛋白的结构。

Eukaryotic post-transcriptional events, such as splicing, transport, and life time regulation of mRNAs, are important for regulation of gene expression and cell differentiation. The splicing of pre-mRNA is one of the important events, which remove the stretches of non-coding RNA before the RNA can be translated into protein. In the early stage in the splicing reaction, 5' splicing site have to be recognized by U1 snRNP,3' splicing site have to be recognized by U2AF protein and the branch site have to be recognized by U2 snRNP. On this study, we investigate the structure of protein which are involved in these components. 1.SURP domain is mainly observed in the splicing machinery (for example U2 snRNP). We determined the structure of SURP domain in SF4 protein. 2.We have tried to reconstitute the p14-SAP155 complex. By NMR method, we confirmed that the complex are successfully reconstituted. 3.In addition, for the structural study of U2AF protein, we could establish the protocol for the reconstitution of U2AF 4.U1C is the one component of the U1 snRNP. In order to gain the information about the interaction for U1-C protein, we have investigated the structure of the N-terminal (1-65) segment of the U1-C protein. And we determined the tertially structure of U1-C (1-61) proteins by homonuclear NMR methods.