ABERRANT RNA PROCESSING IN HUMAN DEVELOPMENT

描述(改编自申请者的描述)：本建议书使用 生物化学和X射线结晶学相结合的方法研究 遗传病中异常RNA加工的结构基础。Cug装订 蛋白质(CUG-BP)、ELAV型RNA结合蛋白(ETR)-1和-3是蛋白质 负责各种发育调控的剪接位点的选择 组织和细胞类型。这些蛋白质含有3个拷贝的rna。 识别基序(RRM)并参与广泛的RNA加工 事件包括剪接、运输和信使核糖核酸周转。Cug-BP、ETR-1和 ETR-3特异性地影响横纹肌组织的剪接位点选择。 Cug-BP首次被分离出来与强直性肌营养不良(DM)有关，这是一种 一种由CUG重复序列扩大引起的遗传病 肌钙蛋白激酶基因的非翻译区，并导致 轻度肌强直到出生时严重发育迟缓。不幸的是，糖尿病是一种常见的 疾病(每8000名新生儿中就有一名)。CuG-BP还与富含CUG的序列相互作用 在许多其他的mRNA中发现。体内CUG-BP、ETR-1水平的变化 1或ETR-3导致组织特异性剪接改变，似乎模仿 发育或疾病相关的RNA加工缺陷。本奖项 将被用来检查已经参与的RNA序列如何 核糖核蛋白复合体被其他RNA结合读取或“扫描” 蛋白质，如那些参与交替剪接的蛋白质。结晶学 确定CUG-BP三维结构所需的条件， ETR-1和ETR-3与其生理相关的RNA靶标通过x- 还将测定射线结晶学。由此得出的结果 这一提议将大大拓宽我们对异常RNA如何 剪接发生并解决RNA新陈代谢的基本方面，以提供 对在人类疾病领域工作的研究人员的即时见解 发病机制。

DESCRIPTION (Adapted from applicant's description): This proposal uses a combination of biochemical and x-ray crystallographic approaches to study the structural basis of aberrant RNA processing in genetic disease. CUG-binding protein (CUG-BP), ELAV-type RNA-binding protein (ETR) -1 and -3 are proteins responsible for developmentally regulated splice site selection in a variety of tissues and cell types. These proteins contain 3 copies of the RNA recognition motif (RRM) and participate in a wide range of RNA processing events including, splicing, transport and mRNA turnover. CUG-BP, ETR-1 and ETR-3 specifically effect splice site selection in striated muscle tissue. CUG-BP was first isolated in connection with myotonic dystrophy (DM), a genetic disease that results from an expansion of CUG repeats in an untranslated region of the myotonin kinase gene and results in symptoms from mild myotonia to severe retardation at birth. DM is an unfortunately common disease (1 in 8000 births). CUG-BP also interacts with CUG-rich sequences found in a number of other mRNAs. Alteration of in vivo levels of CUG-BP, ETR- 1 or ETR-3 result in tissue specific changes in splicing that appear to mimic either developmental or disease related RNA processing defects. This award would be used to examine how RNA sequences already involved in ribonucleoprotein complexes are read or "scanned" by other RNA binding proteins such as those involved in alternate splicing. Crystallographic conditions required to determine the three-dimensional structure of CUG-BP, ETR-1, and ETR-3 bound to their physiologically relevant RNA targets using x- ray crystallography will also be determined. The results derived from this proposal will significantly broaden our understanding of how aberrant RNA splicing occurs and address fundamental aspects of RNA metabolism to provide immediate insights to investigators working in the field of human disease pathogenesis.