The role of post-translational modifications in miRISC function

翻译后修饰在 miRISC 功能中的作用

基本信息

批准号：
9083031
负责人：
John Kim
金额：
$ 33.56万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-25 至 2020-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9083031
关键词：
Address Affect Animal Model Animals Binding Biochemical Biogenesis Biological Biological Process Boxing Caenorhabditis elegans Catabolism Cells Complement Complex Data Development Diagnosis Disease Embryonic Development Ensure Etiology Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Goals Health Heart Diseases Human Human Development Hydroxylation In Vitro Knowledge Lead Malignant Neoplasms Mammals Mediating Messenger RNA Metabolic Diseases Methods MicroRNAs Modeling Molecular Molecular Analysis Muscle Neurons Neuropathy Organogenesis Pathogenesis Pathway interactions Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Play Post-Transcriptional Regulation Post-Translational Protein Processing Prevention Preventive Protein Phosphatase 2A Regulatory Subunit PR53 Proteins Proto-Oncogenes RNA Binding RNA Interference RNA interference screen RNA-Binding Proteins Regulation Regulator Genes Reporting Research Role Specificity Substrate Specificity System Testing Therapeutic Time Tissues Tumor Suppressor Proteins Untranslated RNA base casein kinase II cofactor gene product genome-wide helicase human disease improved in vivo insight mRNA Expression mutant novel novel diagnostics novel therapeutics public health relevance research study therapeutic miRNA therapy design transcriptome

项目摘要

DESCRIPTION (provided by applicant): Post-transcriptional gene silencing mediated by microRNAs (miRNAs) is an established paradigm in metazoan gene regulation, playing critical roles in a wide array of biological processes including developmental timing, organogenesis, and cellular differentiation and proliferation. Mechanisms governing miRNA biogenesis and target silencing are largely established; however, a significant deficit remains in our understanding of how post- translational modifications of the miRNA-mediated silencing complex (miRISC) affect its stability and integrity, subcellular distribution, and effector functio in repressing target mRNA expression. Our long-term goal is to understand how post-translational modifications of miRISC influence miRNA-mediated gene silencing. The specific objective of this proposal is to investigate a novel role for protein kinase CK2 in the miRNA pathway through phosphorylation of core miRISC subunits. Our preliminary data indicate that CK2 physically interacts with miRISC through phosphorylation of two miRISC components: the DEAD-box helicase CGH-1 and the vasa intronic gene product VIG-1. Based on our preliminary data, we hypothesize that through phosphorylation of CGH-1 and VIG-1, CK2 plays a critical role in promoting miRISC binding to target mRNAs. We will test our hypothesis by pursuing two specific aims: 1) Evaluate the key role of CGH-1 and VIG-1 phosphorylation in the specificity of miRNA target binding and expression and their biological significance during animal development. 2) Define the role of CK2 activity and miRISC phosphorylation on the compositional integrity of miRISC and its recruitment to the miRNA target degradation centers, the P bodies. In addition, we will explore potential regulation of CK2 activity by the opposing function of candidate PP2A phosphatases on the phosphorylation state of miRISC. The proposed research is significant because it will establish post-translational modification of miRISC as a general mechanism required for miRISC assembly, localization, and silencing function that impacts all biological pathways regulated by miRNA-mediated gene silencing. The basic insights obtained will also inform the rapidly advancing field of miRNA therapeutics for treating complex human diseases.

描述（由申请人提供）：由 microRNA (miRNA) 介导的转录后基因沉默是后生动物基因调控的既定范例，在发育时序、器官发生以及细胞分化和增殖机制等多种生物过程中发挥着关键作用。控制 miRNA 生物发生和靶标沉默的机制已基本确定；然而，我们对 miRNA 介导的沉默复合物 (miRISC) 的翻译后修饰如何影响其作用的理解仍存在重大缺陷；我们的长期目标是了解 miRISC 的翻译后修饰如何影响 miRNA 介导的基因沉默。我们的初步数据表明，CK2 通过两个 miRISC 成分的磷酸化与 miRISC 发生物理相互作用。 DEAD-box 解旋酶 CGH-1 和 vasa 内含子基因产物 VIG-1 根据我们的初步数据，我们发现通过 CGH-1 和 VIG-1 的磷酸化，CK2 在促进 miRISC 与靶标 mRNA 结合方面发挥着关键作用。我们将通过追求两个具体目标来检验我们的假设：1）评估 CGH-1 和 VIG-1 磷酸化在 miRNA 靶点结合和表达特异性中的关键作用及其生物学意义2) 定义 CK2 活性和 miRISC 磷酸化对 miRISC 组成完整性及其招募到 miRNA 靶降解中心（P 体）的作用。此外，我们将探索候选 PP2A 磷酸酶的相反功能对 CK2 活性的潜在调节。这项研究具有重要意义，因为它将确立 miRISC 的翻译后修饰作为 miRISC 所需的通用机制。组装、定位和沉默功能影响着 miRNA 介导的基因沉默调节的所有生物途径。所获得的基本见解也将为快速发展的 miRNA 治疗领域提供信息，用于治疗复杂的人类疾病。