GENETIC ANALYSIS OF RNA BINDING PROTEINS

描述（改编自申请人摘要）：对RNA结合的兴趣 近年来，随着人们对蛋白质的认识不断提高， 这些蛋白质在各种各样的细胞过程中起着关键作用。 然而，人们对这些结构特征知之甚少。 这些蛋白质使它们能够以如此高的水平识别它们的RNA靶标， 的特异性 这些调查将侧重于查明和 描述了RNA-BP的一些特征，这些特征对它们的生物学行为很重要。 与特定RNA结合的能力。 将特别关注 蛋白质范式代表三种不同的RNA-BP家族： 哺乳动物剪接体蛋白U1 A和U2 B”（RRM家族），HIV-1蛋白 Rev（富含精氨酸的家族）和酵母调节蛋白Mer 1（KH 家庭）。 实验策略将强调遗传 选择/筛选随机突变以鉴定重要的RNA-BP 残基，然后进行定点诱变和生化分析， 更好地了解这些残基在RNA结合中的作用。 的结果 这些研究将提供重要的见解， 不同的RNA-BP家族识别它们的RNA靶点， 阐明了分子生物学的一个关键方面， 明白 这些信息将有助于理解的目标 RNA-BP通常如何能够以高亲和力结合其靶标， 的特异性 鉴于RNA结合蛋白在细胞内的核心作用， HIV-1感染中基因表达的转录后控制， 在脆性X智力迟钝中，这一知识应该是有价值的， 了解发病机制，并最终形成基础， 用于新型基因疗法。

DESCRIPTION (Adapted from applicant's abstract): Interest in RNA-binding proteins (RNA-BPs) has burgeoned in recent years with the growing awareness that these proteins play a key role in a wide variety of cellular processes. However, very little is known about the structural features of these proteins that enable them to recognize their RNA targets with such high specificity. These investigations will focus on identifying and characterizing some of the features of RNA-BPs that are important for their ability to bind to specific RNAs. Particular attention will be devoted to protein paradigms representing three different families of RNA-BPs: the mammalian spliceosomal proteins U1A and U2B" (RRM family), the HIV-1 protein Rev (arginine-rich family), and the yeast regulatory protein Mer1 (KH family). The experimental strategy will emphasize genetic selection/screening of random mutations to identify important RNA-BP residues, followed by site-directed mutagenesis and biochemical analysis to better understand the role of these residues in RNA binding. The results of these studies will provide important insights into how members of three different RNA-BP families recognize their RNA targets, thereby shedding light on a key aspect of molecular biology that presently is poorly understood. This information will contribute to the goal of understanding how RNA-BPs in general are able to bind their targets with high affinity and specificity. In view of the central role that RNA-binding proteins play in the post-transcriptional control of gene expression, in HIV-1 infection, and in fragile X mental retardation, this knowledge should be of value for understanding mechanisms of pathogenesis and could eventually form the basis for novel gene therapies.