Clk Kinases and Splicing Regulation

Clk 激酶和剪接调节

基本信息

批准号：
8471724
负责人：
JOSEPH ADAMS
金额：
$ 28.42万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8471724
关键词：
3&apos Splice Site Address Affect Alternative Splicing Alzheimer&apos s Disease Applications Grants Arginine Ataxia Biological Assay Biological Process C-terminal Cells Data Development Dipeptides Disease Docking Enzymes Event Excision Family Genes Genome Genomics Growth and Development function Investigation Length Link Macromolecular Complexes Malignant Neoplasms Maps Mental disorders Messenger RNA Metabolic Diseases Modeling Muscular Dystrophies N-terminal Names Neurodegenerative Disorders Nuclear Nuclear Protein Parkinsonian Disorders Pattern Phosphorylation Phosphotransferases Play Post-Translational Protein Processing Processed Genes Proline Protein Kinase Protein Splicing Proteins Proteome RNA RNA Splicing Regulation Reporter Genes Role STY kinase Sea Series Serine Site Source Specificity Spliced Genes Spliceosomes Structure Substrate Domain Substrate Specificity Techniques Testing Tissues acrosome stabilizing factor base human disease inorganic phosphate novel protein function prototype seryl-proline

项目摘要

DESCRIPTION (provided by applicant): While the proper selection of splice sites is essential for genomic diversity, adaptive growth and development, errors in splicing can have enormous detrimental effects on function and are 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 disease. 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' and 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 understandig of how these different forms are attained is, at best, incomplete. SR protein nuclear entry and splicing function are driven by a basal level of phosphorylation (hypo-phosphorylation) catalyzed by the SRPK family of protein kinases. However, this simple paradigm of one kinase-one substrate is now being challenged as a second protein kinase family has emerged as critical SR protein regulators. The Clk family of kinases can increase SR protein phosphoryl content to a greater extent than the SRPKs, generating hyper-phosphorylated forms that are largely uncharacterized at both structural and functional levels. While they differ substantially from the SRPKs in several ways, most importantly, Clk kinases possess an additional noncatalytic domain that we showed recently is responsible for its unique hyper-phosphorylating activity. Despite its significance in controlling SR protein function and splicing, little is known about the Clk enzymes. Using novel phosphate mapping and structural techniques combined with cell-based assays, we will investigate how the Clk kinases hyper-phosphorylate SR proteins and modulate splicing.

描述（由申请人提供）：虽然剪接位点的正确选择对于基因组多样性，适应性生长和发育至关重要，但剪接错误可能会对功能产生巨大的有害影响，现在被认为是许多人类疾病的根本原因。实际上，剪接错误与肌肉营养不良，阿尔茨海默氏病，帕金森氏症，精神疾病，共济失调和癌症有关，从而研究了控制剪接位点选择对人类疾病至关重要的因素。剪接发生在剪接体，这是一种由多种RNA和许多蛋白质组成的大分子复合物。对正常基因剪接至关重要的是正确选择5'和3'剪接位点，事件发生在剪接体的发展初期，其特异性是由一种称为SR蛋白的剪接因子家族的指导。 SR蛋白的磷酸化状态直接影响其亚细胞定位和剪接活动，但我们对如何获得这些不同形式的理解充其量是不完整的。 SR蛋白核进入和剪接功能是由蛋白激酶SRPK家族催化的基础磷酸化（低磷酸化）驱动的。然而，随着第二个蛋白激酶家族的质疑，这种简单的一种激酶 - 一个底物现在受到挑战。激酶的clk家族可以比SRPK更大程度地增加SR蛋白磷酸含量，从而产生高磷酸化的形式，这些形式在结构和功能水平上都在很大程度上没有表征。尽管它们在几种方面与SRPK有很大差异，但最重要的是，CLK激酶具有额外的非催化域，我们最近显示的是其独特的高磷酸化活性。尽管它在控制SR蛋白功能和剪接方面具有重要意义，但对 CLK酶。使用新型的磷酸盐映射和结构技术与基于细胞的测定结合，我们将研究CLK激酶如何高磷酸化的SR蛋白和调节剪接。