Mechanism for Activation of the Antiviral Kinase PKR

抗病毒激酶 PKR 的激活机制

基本信息

批准号：
8588247
负责人：
JAMES L COLE
金额：
$ 39.22万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-12-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8588247
关键词：
Adenoviruses Adopted Adverse effects Affect Affinity Antiviral Agents Antiviral Therapy Binding Binding Sites Biological Biological Assay Biological Models C-terminal Cell Proliferation Complex Dependence Development Digestion Dimerization Double-Stranded RNA Enzymes Eukaryotic Initiation Factors Foundations Funding Growth Factor HIV Heparin Holoenzymes Immune System Diseases Immune response Interferons Length Libraries Link Macromolecular Complexes Measures Mediating Metabolic Diseases Metabolism Methods Modeling Molecular Molecular Conformation N-terminal Natural Immunity Nerve Degeneration Neutrons Nutrient Oligosaccharides Orthologous Gene Pathway interactions Peptide Hydrolases Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Protein Biosynthesis RNA RNA Binding RNA Editing RNA Interference Reaction Regulation Research Resistance Roentgen Rays Role Signal Transduction Small RNA Stress Structural Models Structure Structure-Activity Relationship Tail Theoretical model Therapeutic Agents VAI RNA Viral Viral Cancer Viral Proteins Virus Diseases Withdrawal Work base biological adaptation to stress biophysical techniques cancer therapy cell growth cytokine design dimer eIF-2 Kinase flexibility high throughput analysis in vivo inhibitor/antagonist monomer neoplastic novel programs resistance mechanism response sedimentation equilibrium sedimentation velocity small molecule stem therapeutic effectiveness viral resistance

项目摘要

DESCRIPTION (provided by applicant): The RNA-activated protein kinase, PKR, is induced by interferon and plays a pivotal role in the innate immunity response to viral infection. PKR is synthesized in a latent state, but upon binding activating RNAs it undergoes autophosphorylation reactions that activate the kinase. Phosphorylation of the eukaryotic initiation factor eIF2a by PKR inhibits viral protein synthesis. PKR is also activated by cytokines, growth factor withdrawal and stress signals and participates in pathways regulating stress response, cellular growth and proliferation, nutrient signaling and metabolism. The broad objective of our research program is to define the molecular mechanisms that regulate the activity of PKR using quantitative biophysical and structural methods. In Aim I we will define the interactions of PKR with complex RNA inhibitors and activators. Specifically, we will identify the structural features within the Adenovirus VAI RNA that mediate high-affinity PKR binding without activating the kinase, determine the mechanism of activation of PKR by novel single stranded-double stranded RNAs and correlate PKR dimerization and activation with the length and flexibility of a linker region in model RNAs. PKR represents an attractive target for development of novel antiviral therapies to enhance the innate immune response and in Aim 2 we will use a nonradioactive assay to screen libraries of drug-like compounds for novel PKR activators. In Aim 3, small angle scattering methods will be used to determine the conformation of ordered forms of PKR and to develop structural model of complexes of PKR with RNA inhibitors and activators. These studies will provide the foundation for the design of therapeutic agents that target PKR for the treatment of viral infections and cancer.

描述（由申请人提供）：RNA激活的蛋白激酶PKR由干扰素诱导，并在对病毒感染的先天免疫反应中起关键作用。 PKR是在潜在状态下合成的，但是结合激活RNA后，它会经历激活激酶的自磷酸化反应。 PKR抑制病毒蛋白合成的真核起始因子EIF2A的磷酸化。 PKR还被细胞因子，生长因子戒断和应激信号激活，并参与调节压力反应，细胞生长和增殖，养分信号传导和代谢的途径。我们研究计划的广泛目标是定义使用定量生物物理和结构方法来调节PKR活性的分子机制。在目的中，我们将定义PKR与复杂的RNA抑制剂和激活剂的相互作用。具体而言，我们将确定腺病毒VAI RNA中介导高亲和力PKR结合而无需激活激酶的结构特征，通过新型的单链双链RNAS确定PKR激活的机制，并与模型RNAS中接头区域的长度和灵活性相关的PKR PKR Dimerization和激活。 PKR代表了开发新型抗病毒疗法以增强先天免疫反应的有吸引力的靶标，在AIM 2中，我们将使用非放射活性测定法对新型PKR激活剂进行类似药物的化合物的库。在AIM 3中，小角度散射方法将用于确定PKR有序形式的构象，并开发PKR与RNA抑制剂和激活剂的结构模型。这些研究将为靶向PKR的治疗剂治疗病毒感染和癌症的设计奠定基础。