ALTERNATIVE SPLICING REGULATION AND MEMBRANE TRAFFICKING

选择性剪接调节和膜运输

基本信息

批准号：
10569039
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
项目类别：
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/10569039
关键词：
Adult Affect Alternative Splicing Animals Architecture Binding Binding Sites Birth Brain 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 Future Gene Expression Genes Genetic Materials Health Heart Diseases Histone Acetylation Histone Deacetylase Inhibitor Human Individual Institution Intracellular Transport Kinetics Knowledge 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 cardiogenesis 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 posttranscriptional programs protein function protein transport trafficking

项目摘要

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/总结生物体的复杂性与蛋白质编码基因的数量无关。监管机制有助于进化过程中基因功能的多样化。选择性拼接是转录后机制，解释单个基因如何产生多个转录本包含或排除特定区域。在人类中，超过 90% 的基因经历选择性剪接，与高等真核生物的细胞和功能复杂性增加相一致。全基因组研究已具有完全未知功能的剪接异构体和网络的数量呈指数增加。基因编码膜运输蛋白在发育过程中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意义的问题这一级别的监管。了解剪接调节在蛋白质表达和功能中的作用参与贩运和膜动力学是启发我们项目的知识差距。 R01提案的科学前提是运输蛋白的选择性剪接调控在细胞、组织和器官的发育中发挥着关键的作用。该提案提出的基本问题这就是选择性剪接如何控制特定组织和细胞类型中的膜运输。我们将解决这个问题问题有两个目标：（目标 1）协调这些剪接转变的调控机制是什么？（目标 2）膜运输基因剪接调节的功能后果是什么？在具体目标 1 中，我们将确定两种 RNA 结合蛋白（PTBP 和 QK）的作用和表观遗传学肌肉细胞分化中膜运输基因的剪接调节。在具体目标 2 中，我们将确定膜运输的选择性剪接调节的下游功能后果基因 Trip10（Cdc42 相互作用蛋白 4，CIP4）利用细胞培养实验和动物研究。全面的，该项目完成后，我们将确定选择性剪接所涉及的分子机制膜运输蛋白的调节及其生理意义。