Clk Kinases and Splicing Regulation

Clk 激酶和剪接调节

• 批准号: 8294209
• 负责人: JOSEPH ADAMS
• 金额: $ 29.44万
• 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 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8294209
• 关键词: 3' Splice Site; Address; Affect; Alternative Splicing; Alzheimer' 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

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. PUBLIC HEALTH RELEVANCE: Although many human diseases including neurodegenerative disorders and cancers are linked to mistakes in gene splicing, what controls the selection of splice sites in the cell is still poorly understood. To understand the link between disease and gene processing, we are studying how unique phosphorylation states in an essential group of splicing factors (SR proteins) are catalyzed by the Clk family of enzymes and regulate the selection of correct splice sites.

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、适应性生长和发育是必不可少的，但剪接中的错误可能对功能具有巨大的有害影响，并且现在被认为是许多人类疾病的根本原因。事实上，剪接错误与肌肉萎缩症、阿尔茨海默病、帕金森综合征、精神疾病、共济失调和癌症有关，使得控制剪接位点选择的因素的研究对人类疾病至关重要。剪接发生在剪接体上，剪接体是由几种RNA和许多蛋白质组成的大分子复合物。正常基因剪接的关键是正确选择5'和3'剪接位点， 发生在剪接体发育的早期，其特异性由称为SR蛋白的剪接因子的基本家族指导。SR蛋白的磷酸化状态直接影响它们的亚细胞定位和剪接活性，但我们对这些不同形式是如何获得的理解充其量是不完整的。SR蛋白核进入和剪接功能由SRPK蛋白激酶家族催化的基础水平的磷酸化（低磷酸化）驱动。然而，这种一种激酶-一种底物的简单范例现在正受到挑战，因为第二种蛋白激酶家族已经成为关键的SR蛋白调节剂。激酶的Clk家族可以比SRPK更大程度地增加SR蛋白磷酰基含量，产生在结构和功能水平上都很大程度上未表征的超磷酸化形式。虽然它们在几个方面与SRPKs有很大的不同，但最重要的是，Clk激酶拥有一个额外的非催化结构域，我们最近发现这是其独特的超磷酸化活性的原因。尽管它在控制SR蛋白功能和剪接方面具有重要意义，但人们对SR蛋白的功能知之甚少。 Clk酶。使用新的磷酸映射和结构技术结合细胞为基础的测定，我们将研究如何的Clk激酶超磷酸化SR蛋白和调节剪接。 公共卫生相关性：尽管许多人类疾病，包括神经退行性疾病和癌症，都与基因剪接中的错误有关，但控制细胞中剪接位点选择的因素仍然知之甚少。为了了解疾病和基因加工之间的联系，我们正在研究如何独特的磷酸化状态的剪接因子（SR蛋白）的一个重要的组被催化的Clk家族的酶和调节正确的剪接位点的选择。