Coordination of SR Protein Phosphorylation and RNA Splicing

SR 蛋白磷酸化和 RNA 剪接的协调

• 批准号: 7990450
• 负责人: JOSEPH ADAMS
• 金额: $ 29.3万
• 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 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7990450
• 关键词: 3' Splice Site; Address; Affinity; Alternative Splicing; Arginine; Ataxia; Biological Assay; Biological Process; C-terminal; Cell Nucleus; Cells; Cellular Assay; Chemicals; Cisplatin; Communities; Complex; Cytoplasm; Dipeptides; Disease; Docking; Down-Regulation; Drug Delivery Systems; Engineering; Enzymes; Event; Family; Funding; Genes; Goals; Health; Hereditary Disease; Human; Human Genome; Kinetics; Lead; Link; Lower Organism; Malignant Neoplasms; Mental disorders; Messenger RNA; Methods; Modification; Molecular Conformation; Muscular Dystrophies; Mutation; N-terminal; Names; Neurodegenerative Disorders; Neurofibromatoses; Organelles; Pancreatic carcinoma; Parkinsonian Disorders; Pathway interactions; Pattern; Phosphorylation; Phosphorylation Site; Phosphotransferases; Protein Binding; Protein Conformation; Protein Family; Protein Kinase; Protein Splicing; Proteins; RNA; RNA Processing; RNA Recognition Motif; RNA Splicing; Reaction; Refractory; Retinoblastoma; Role; Serine; Site; Specificity; Spliced Genes; Spliceosome Assembly Pathway; Spliceosomes; Structure; Testicular Neoplasms; Testing; acrosome stabilizing factor; chemotherapeutic agent; chemotherapy; human disease; interest; mRNA Precursor; macromolecular assembly; novel; protein function; protein structure; prototype; public health relevance; treatment strategy; tumor

DESCRIPTION (provided by applicant): Comparisons of the genomes from humans and lower organisms reveal that the complexity in humans is achieved not by a dramatic increase in the number of genes but by alternative splicing events that stitch together different portions of genes to generate diverse proteins. Correct splicing allows normal healthy function, however, incorrect splicing is linked to many human diseases. For example, muscular dystrophy, ataxias, parkinsonism, neurofibromatosis, psychiatric disorders and cancer have their origins in splicing errors. Splicing reactions are catalyzed by a large macromolecular machine known as the spliceosome. Composed of both RNA and protein, the spliceosome can accurately select the proper splice sites from a considerably large precursor mRNA in healthy cells. The assembly of the spliceosome, the identification of the correct 5' and 3' splice sites and the chemical splicing reaction itself is regulated by a large class of splicing factors known as SR proteins. SR proteins contain one or two RNA recognition motifs and a long C-terminal domain rich in numerous arginine-serine dipeptide repeats. The phosphorylation of the RS domain serves many RNA processing functions including splice-site selection, import of SR proteins into the nucleus and export of mature mRNA to the cytoplasm. This project will investigate how two principal families of splicing enzymes uniquely impact SR protein function through regiospecific, multi-site phosphorylation of the RS domains. Using engineered footprinting methods, the directionality of the splicing enzymes will be defined and shown to control which serines in the RS domain are modified. The effects of these selective phosphorylation reactions on SR protein structure and interaction/function within the spliceosome will then be evaluated using kinetic, structural, splicing and cellular assays. The goal is to identify how splicing kinases recognize and phosphorylate specific regions of the RS domains and determine how these chemical modifications impact splicing componentry. PUBLIC HEALTH RELEVANCE: Health Relevance Statement Comparisons of the genomes from humans and lower organisms reveal that the complexity in humans is achieved not by a dramatic increase in the number genes but by splicing events that stitch together different portions of genes to generate diverse proteins. Correct splicing allows normal healthy function, however, incorrect splicing is linked to many human neurodegenerative diseases and cancer. We are investigating how the newly identified drug targets for diverse diseases known as splicing enzymes (named SR-kinases) regulate important splicing factors (a specific family of proteins known as SR proteins) which cooperate in the control of alternative splicing reactions important in both health and disease.

描述（由申请人提供）：人类和低等生物基因组的比较表明，人类的复杂性不是通过基因数量的急剧增加来实现的，而是通过将基因的不同部分缝合在一起以产生不同蛋白质的选择性剪接事件来实现的。正确的剪接可以保证正常的健康功能，然而，不正确的剪接与许多人类疾病有关。例如，肌肉营养不良、共济失调、帕金森病、神经纤维瘤病、精神疾病和癌症都起源于剪接错误。剪接反应由称为剪接体的大分子机器催化。剪接体由RNA和蛋白质组成，可以从健康细胞中相当大的前体mRNA中准确地选择合适的剪接位点。剪接体的组装、正确 5' 和 3' 剪接位点的识别以及化学剪接反应本身受到一大类称为 SR 蛋白的剪接因子的调节。 SR 蛋白含有一或两个 RNA 识别基序和一个富含大量精氨酸-丝氨酸二肽重复序列的长 C 端结构域。 RS 结构域的磷酸化具有许多 RNA 加工功能，包括剪接位点选择、将 SR 蛋白导入细胞核以及将成熟 mRNA 输出到细胞质。该项目将研究两个主要剪接酶家族如何通过 RS 结构域的区域特异性、多位点磷酸化来独特地影响 SR 蛋白功能。使用工程足迹法，将定义剪接酶的方向性并显示其控制 RS 结构域中的丝氨酸被修饰。然后将使用动力学、结构、剪接和细胞测定来评估这些选择性磷酸化反应对剪接体内SR蛋白结构和相互作用/功能的影响。目标是确定剪接激酶如何识别和磷酸化 RS 结构域的特定区域，并确定这些化学修饰如何影响剪接成分。公共卫生相关性：健康相关性声明 人类和低等生物基因组的比较表明，人类的复杂性不是通过基因数量的急剧增加来实现的，而是通过将基因的不同部分缝合在一起以产生不同蛋白质的剪接事件来实现的。正确的剪接可以实现正常的健康功能，然而，不正确的剪接与许多人类神经退行性疾病和癌症有关。我们正在研究新发现的针对多种疾病的药物靶点，称为剪接酶（称为 SR 激酶），如何调节重要的剪接因子（称为 SR 蛋白的特定蛋白质家族），这些因子在控制对健康和疾病都很重要的选择性剪接反应中合作。