Regulatory Pathways of SR Protein Kinases

SR蛋白激酶的调控途径

• 批准号: 9235874
• 负责人: JOSEPH ADAMS
• 金额: $ 31.78万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9235874
• 关键词: Address; Affect; Affinity; Alzheimer' s Disease; Arginine; Ataxia; Attention; Biological Assay; C-terminal; Cardiovascular Diseases; Carrier Proteins; Catalysis; Cell Nucleus; Cell physiology; Cells; Complex; Cytoplasm; Disease; EGF gene; Enzymatic Biochemistry; Enzymes; Excision; Family; Funding; Genes; Growth Factor; Human; Kinetics; Knowledge; Lead; Link; Macromolecular Complexes; Malignant Neoplasms; Messenger RNA; Molecular Chaperones; Muscular Dystrophies; Nature; Normal Cell; Nuclear; Nuclear Import; Nuclear Protein; Organism; Parkinsonian Disorders; Peptide Hydrolases; Phosphorylation; Phosphotransferases; Protein Family; Protein Kinase; Protein Splicing; Proteins; Proteome; RNA; RNA Splicing; Reaction; Regulation; Regulatory Pathway; Research Project Grants; Role; Serine; Site; Spliced Genes; Spliceosomes; Techniques; U1 Small Nuclear Ribonucleoprotein; catalyst; cellular imaging; genome-wide; human disease; inorganic phosphate; mRNA Precursor; mutant; novel; nucleocytoplasmic transport; pressure; protein transport; public health relevance; release factor

Project Summary/Abstract: Splicing converts a single gene into multiple unique mRNA fragments to expand the size of the proteome and regulate cell function. While splicing is integral for normal function in complex organisms, mistakes in splice-selection can lead to disease. In fact, splicing errors are associated with numerous human disorders including muscular dystrophy, Alzheimer's disease, parkinsonism, cardiovascular disease, ataxias and cancers. Splicing occurs at the spliceosome, a macromolecular complex that includes both RNA and protein. In the latter group, SR proteins are essential splicing factors that control where the spliceosome assembles on precursor mRNA. SR proteins contain C-terminal domains rich in arginine-serine repeats whose polyphosphorylation controls splice-site selection. The SRPK family of protein kinases phosphorylates these RS domains directing SR proteins into the nucleus for splicing activity. While SRPKs are normally localized to the cytoplasm for this function, they can enter the nucleus under certain conditions but their function in this cellular compartment is not well understood. We will now investigate the functions of SRPKs in both the cytoplasm and nucleus using genome-wide splicing assays, cell imaging, fast-mixing kinetics, protease footprinting and structural techniques. We will determine how the unique SRPK phosphorylation mechanism governs transport of SR proteins from the cytoplasm to the nucleus. We will explore factors that regulate SRPK nuclear import through a novel kinase- kinase complex. Finally, we will investigate how this complex affects splicing reactions through the mobilization of SR proteins in the nucleus. Overall, the studies outlined in this proposal will greatly expand our knowledge of SRPK-induced phosphorylation of SR proteins and their splicing function.

项目摘要/摘要： 剪接将单个基因转换为多个独特的mRNA片段以扩展 蛋白质组的大小和调节细胞功能。而拼接对于正常功能不可或缺 复杂的生物体，剪接选择中的错误会导致疾病。实际上，拼接错误 与许多人类疾病有关，包括肌肉营养不良，阿尔茨海默氏症 疾病，帕金森氏症，心血管疾病，共济失调和癌症。剪接发生在 剪接体是一种大分子复合物，包括RNA和蛋白质。在后者 组，SR蛋白是控制剪接体组装的基本剪接因子 在前体mRNA上。 SR蛋白包含富含精氨酸丝氨酸重复序列的C末端结构域 多磷酸化控制剪接位点的选择。 SRPK蛋白激酶家族 磷酸化这些RS结构域，将SR蛋白引导到细胞核中以进行剪接活性。 尽管SRPK通常位于此功能的细胞质中，但它们可以进入 核在某些条件下，但它们在该细胞室中的功能不好 理解。现在，我们将研究SRPK在细胞质和 使用全基因组剪接测定，细胞成像，快速混合动力学，蛋白酶的细胞核 足迹和结构技术。我们将确定唯一的srpk 磷酸化机制控制SR蛋白从细胞质转运到 核。我们将探索通过新型激酶 - 调节SRPK核进口的因素 激酶复合物。最后，我们将研究这种复合物如何通过 核中SR蛋白的动员。总体而言，该提案中概述的研究将 大大扩展了我们对SRPK诱导的SR蛋白磷酸化及其知识 剪接功能。