Ribonucleoprotein Complexes Regulating T-Cell Activation

描述（由申请人提供）：该提案概述了一个计划，结合基因组数据分析，生物化学，结构生物学和功能表征，以了解核糖核蛋白复合物在调节t细胞活化中的作用。表达数据被挖掘，以确定一组rna结合蛋白，这些蛋白在t细胞激活时要么差异表达，要么选择性剪接。这些rna结合蛋白构成了结构基因组学工作的输入，代表了三个结构靶流中的第一个。这些rna结合蛋白的蛋白相互作用伙伴已经从Human Interactome数据库中被鉴定出来，特异性蛋白-蛋白复合物的形成将在t细胞和体外得到证实。产生的蛋白质-蛋白质复合物将作为第二个目标流输入到结构基因组学工作中。RNA结合蛋白的RNA靶点将使用高通量测序和交联（HITS-CLIP）进行鉴定，并在t细胞和体外进行确认。由此产生的rna -蛋白复合物将作为第三个目标流输入到结构基因组学工作中。对于结构基因组学研究中出现的蛋白质、蛋白质-蛋白质复合物和rna -蛋白质复合物，将在t细胞中进行功能验证，以确定这些特定成分在t细胞活化中的作用。这项工作将为研究这种结构提供重要的新见解

DESCRIPTION (provided by applicant): This proposal outlines a plan to combine genomic data analysis, biochemistry, structural biology, and functional characterization to understand the role of ribonucleoprotein complexes in regulating T-cell activation. Expression data were mined to identify a set of RNA-binding proteins that were either differentially expressed or alternatively spliced, upon T-cell activation. These RNA-binding proteins constitute the input into a structural genomics effort, representing the first of three structural target streams. Protein interacting partners for these RNA-binding proteins have been identified from the Human Interactome Database, and the formation of specific protein-protein complexes will be confirmed both in T-cells and in vitro. The resulting set of protein-protein complexes will be input into a structural genomics effort as the second target stream. The RNA targets for the RNA-binding proteins will be identified using high throughput sequencing and crosslinking (HITS-CLIP), and confirmed in T-cells and in vitro. The resulting set of RNA-protein complexes will be input into a structural genomics effort as the third target stream. For the proteins, protein-protein complexes, and RNA-protein complexes that emerge from the structural genomics effort, functional validation will be carried out in T-cells to determine the role of these specific components in T-cell activation. This work should provide significant new insights into the structure of ribonucleoprotein complexes in general, as well as insights into how these complexes are involved in posttranscriptional gene regulation.