Clk Kinases and Splicing Regulation

Clk 激酶和剪接调节

• 批准号: 9177458
• 负责人: JOSEPH ADAMS
• 金额: $ 32.65万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9177458
• 关键词: 3' Splice Site; Active Sites; Affect; Alzheimer' s Disease; Ataxia; Binding; Biochemical; Biological; Biological Assay; Biological Process; C-terminal; Cell Nucleus; Cell Survival; Cell physiology; Cells; Chemicals; Complex; Data; Development; Dipeptides; Disease; Docking; Enzymes; Event; Family; Funding; Genes; Genomics; Goals; Growth and Development function; Health; Human; Image; In Vitro; Life; Link; Macromolecular Complexes; Malignant Neoplasms; Maps; Mediator of activation protein; Mental disorders; Messenger RNA; Methods; Modification; Molecular Conformation; Muscular Dystrophies; N-terminal; Nature; Nuclear; Parkinsonian Disorders; Pattern; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Protein Conformation; Protein Family; Protein Kinase; Protein phosphatase; Proteins; RNA Splicing; RRM1 gene; RRM2 gene; Regulation; Role; STY kinase; Site; Specificity; Spliced Genes; Spliceosomes; Structure; U1 Small Nuclear Ribonucleoprotein; abstracting; acrosome stabilizing factor; biophysical techniques; genome-wide; human disease; in vivo; public health relevance; research study; scaffold; seryl-proline

Project Summary/Abstract: While the proper selection of splice sites drives genomic diversity, adaptive growth and development, errors in splicing can have enormous detrimental effects on function and is now recognized as the underlying cause for many human diseases. Indeed, splicing errors are associated with muscular dystrophy, Alzheimer's disease, Parkinsonism, psychiatric disorders, ataxias and cancers making the study of factors that control splice-site selection vitally important for human health. Splicing occurs at the spliceosome, a macromolecular complex composed of several RNAs and numerous proteins. Critical to normal gene splicing is the proper selection of the 5'-3' splice sites, events that occur early in the development of the spliceosome and whose specificity is guided by an essential family of splicing factors known as SR proteins. The phosphorylation states of SR proteins directly impact their subcellular localization and splicing activities but our understanding of how these different forms are attained is, at best, incomplete. The CLK family of protein kinases phosphorylates SR proteins and mobilizes them to sites of active gene splicing. The CLKs differ from many classic protein kinases in that they lack a docking groove for substrate binding but instead contain a disordered N-terminal extension that we showed attaches to the SR protein. In this proposal we will explore the role of the N-terminus for the mobilization of CLK1 and recognition of SR proteins in the nucleus using a wide array of in vivo and in vitro experiments. We will study the effects of CLK-dependent phosphorylation on SR protein conformation, subcellular localization, and interactions with critical mediators of splice-site selection in the spliceosome. The larger goal of this proposal is to define CLK function at both biological and biochemical levels so that we can better understand the mechanisms of human diseases associated with errors in splicing.

项目摘要/摘要： 而正确选择的剪接站点可以驱动基因组多样性，适应性增长和 开发，剪接错误可能会对功能产生巨大的有害影响，现在是 被认为是许多人类疾病的根本原因。确实，剪接错误是关联的 有肌肉营养不良，阿尔茨海默氏病，帕金森氏症，精神疾病，共济失调和 对控制剪接站点选择的因素进行研究对人类健康至关重要。 剪接发生在剪接体，这是一种大分子复合物，由几个RNA和 许多蛋白质。对正常基因剪接至关重要的是正确选择5'-3'剪接位点， 剪接体发展早期发生的事件，其特异性由 剪接因子的基本家族称为SR蛋白。 SR蛋白的磷酸化态 直接影响其亚细胞本地化和剪接活动，但我们对这些活动的理解 充其量是不完整的。蛋白激酶的CLK家族磷酸化 SR蛋白并将它们动员到活性基因剪接位置。库与许多经典不同 蛋白激酶因为它们缺乏用于底物结合的对接凹槽，但含有无序 我们显示的N末端延伸是附着在SR蛋白上的。在此提案中，我们将探索 N末端在动员CLK1和使用SR蛋白识别核中的作用 各种体内和体外实验。我们将研究CLK依赖性的影响 SR蛋白构象，亚细胞定位以及与关键的相互作用的磷酸化 剪接位置选择剪接位点的介体。该提议的更大目标是定义clk 在生物学和生化水平上的功能，以便我们可以更好地了解 与剪接错误相关的人类疾病。