ALTERNATIVE SPLICING REGULATION AND MEMBRANE TRAFFICKING

选择性剪接调节和膜运输

• 批准号: 10360604
• 负责人: Jimena Giudice
• 金额: $ 32.1万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10360604
• 关键词: Adult; Affect; Alternative Splicing; Animals; Architecture; Binding; Binding Sites; Birth; Brain; Cardiac; Cell Culture Techniques; Cell Differentiation process; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Confocal Microscopy; Coupling; Cytoskeleton; DNA; Development; Doctor of Philosophy; Elements; Environment; Epigenetic Process; Eukaryota; Event; Evolution; Exclusion; Exons; Faculty; Fetal Development; Future; Gene Expression; Genes; Genetic Materials; Health; Heart; Histone Deacetylase Inhibitor; Histones; Human; Individual; Intracellular Transport; Kinetics; Knowledge; Lead; Life; Maintenance; Malignant Neoplasms; Measures; Membrane; Mentors; Molecular; Mus; Muscle; Muscle Cells; Muscle Development; Muscle Fibers; Muscular Dystrophies; Mutate; Myoblasts; Myocardium; Myopathy; Neonatal; Neurons; Organ; Organism; Output; Pathologic; Pattern; Physiological; Physiology; Play; Positioning Attribute; Postdoctoral Fellow; Process; Protein Isoforms; Proteins; RNA; RNA Polymerase II; RNA Splicing; RNA-Binding Proteins; Regulation; Research; Research Personnel; Role; Skeletal Muscle; Striated Muscles; TRIP10 gene; Tertiary Protein Structure; Time; Tissue-Specific Splicing; Tissues; Training; Transcript; cell type; disease-causing mutation; exon skipping; experimental study; fetal; flexor digitorum brevis; gene function; genome-wide analysis; human disease; in vivo; link protein; magnetic beads; mechanical properties; mouse model; nervous system disorder; postnatal; postnatal development; programs; protein function; protein transport; trafficking

ABSTRACT / SUMMARY The complexity of organisms does not correlate with the number of protein encoding genes. Regulatory mechanisms have contributed to the diversification of gene function during evolution. Alternative splicing is a posttranscriptional mechanism that explains how single genes can produce more than one transcript due to the inclusion or exclusion of specific regions. In humans, more than 90% of genes undergo alternative splicing, consistent with the increased cellular and functional complexity of higher eukaryotes. Genome wide studies have exponentially increased the number of splicing isoforms and networks with completely unknown functions. Genes encoding membrane trafficking proteins are developmentally regulated by alternative splicing specifically in striated muscles between birth and adulthood. This finding raises the question of the physiological implications of this level of regulation. Understanding the role of splicing regulation in the expression and function of proteins involved in trafficking and membrane dynamics is the knowledge gap inspiring our project. The scientific premise of this R01 proposal is that alternative splicing regulation of trafficking proteins plays key developmental roles in cells, tissues, and organs. The fundamental question asked in this proposal is how alternative splicing controls membrane trafficking in specific tissues and cell types. We will tackle this question in two aims: (aim 1) what are the regulatory mechanisms that coordinate these splicing transitions? (aim 2) what are the functional consequences of splicing regulation of membrane trafficking genes? In Specific Aim 1, we will identify the role of two RNA-binding proteins (PTBP and QK) and epigenetics in splicing regulation of membrane trafficking genes in muscle cell differentiation. In Specific Aim 2, we will determine the downstream functional consequences of alternative splicing regulation of the membrane trafficking gene Trip10 (Cdc42 interacting protein-4, CIP4) utilizing cell culture experiments and animal studies. Overall, after completion of this project we will have identified the molecular mechanisms involved in alternative splicing regulation of membrane trafficking proteins, and their physiological significance.

摘要/摘要 生物体的复杂性与编码基因的蛋白质的数量无关。监管 在进化过程中，机制促成了基因功能的多样化。可选剪接是一种 转录后机制，解释单个基因如何能够产生多个转录本 包括或排除特定地区。在人类中，超过90%的基因经历了选择性剪接， 与高等真核生物细胞和功能复杂性的增加相一致。全基因组研究已经 具有完全未知功能的剪接异构体和网络的数量呈指数级增加。基因 编码膜运输蛋白的发育受到选择性剪接的调节，特别是在 出生到成年之间的横纹肌。这一发现提出了生理学含义的问题 这一级别的监管。理解剪接调控在蛋白质表达和功能中的作用 参与贩运和膜动力学是激励我们项目的知识鸿沟。 R01提案的科学前提是运输蛋白的选择性剪接调控 在细胞、组织和器官中起着关键的发育作用。这项提案提出的根本问题是 选择性剪接是如何控制特定组织和细胞类型中的膜运输的。我们会解决这个问题的 两个目标中的问题：(目标1)协调这些剪接过渡的调控机制是什么？ (目的2)膜转运基因剪接调控的功能后果是什么？ 在具体目标1中，我们将确定两个RNA结合蛋白(PTBP和QK)和表观遗传学的作用 在肌肉细胞分化中膜运输基因的剪接调控。在具体目标2中，我们将 确定膜转运的选择性剪接调控的下游功能后果 Trip10基因(CDC42相互作用蛋白-4，CIP4)利用细胞培养实验和动物实验。总的来说， 在这个项目完成后，我们将确定参与选择性剪接的分子机制。 膜转运蛋白的调控及其生理意义。