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：
7655726
负责人：
JAMES L COLE
金额：
$ 29.97万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7655726
关键词：
Adverse effects Affect Affinity Antiviral Agents Antiviral Response Apoptosis Binding Biological Biological Assay Cell physiology Cells Coupled Coupling Dimerization Double-Stranded RNA Effectiveness Equilibrium Eukaryotic Initiation Factors Foundations Free Energy Funding Heparin Hepatitis C virus Immune System Diseases Influenza B virus Interferons Macromolecular Complexes Malignant Neoplasms Mediating Methods Modeling Molecular Mutation Natural Immunity Oligosaccharides PTPN11 gene Pathway interactions Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Population Prokaryotic Initiation Factor-2 Protein Array Protein Biosynthesis Protein Synthesis Inhibition Proteins Public Health RNA RNA Interference Pathway Reaction Regulation Research Resistance Role Solutions Structure-Activity Relationship Testing Therapeutic Agents Therapeutic Intervention Treatment Protocols Viral Virulence Virus Virus Diseases base combat design eIF-2 Kinase influenza virus INS1 protein influenzavirus inhibitor/antagonist mortality neoplastic novel pandemic disease programs resistance mechanism response small molecule stoichiometry therapeutic effectiveness viral resistance

项目摘要

The double-stranded (ds) RNA-activated protein kinase, PKR, is one of several proteins induced by interferon and plays a pivotal role in the cellular antiviral response. PKR has also been implicated in other cellular processes including transformation, differentiation and apoptosis. There are also structural and functional connections between PKR and the RNA interference (RNAi) pathway. PKR is synthesized in a latent state and is activated upon binding to dsRNA to undergo autophosphorylation reactions that activate the kinase. In turn, activated PKR phosphorylates eukaryotic initiation factor 2a, resulting in the inhibition of protein synthesis in virally-infected cells. The importance of this antiviral pathway is highlighted by the diverse mechanisms that viruses have evolved to combat PKR. The broad objective of our research program is to define the molecular mechanisms for activation and inhibition of PKR. We will define the stoichiometries, affinities and free-energy coupling that govern formation of macromolecular complexes that regulate PKR activity using quantitative biophysical and structural methods. We will determine how NS5A from hepatitis C virus and NS1 from influenza virus interact with PKR to evade the antiviral pathway. Mutations in these proteins that affect virulence or confer interferon resistance will be correlated with PKR binding and inhibition. We will determine how short, heparin oligosaccharides function as PKR activators. Microarrays of synthetic oligosaccharides will be screened and structure-activity relationships will be generated to define novel small-molecule activators of PKR. These studies will provide the foundation for the design of therapeutic agents that target PKR for the treatment of viral infections and cancer.

双链（DS）RNA激活的蛋白激酶PKR是干扰素诱导的几种蛋白质之一，在细胞抗病毒反应中起关键作用。 PKR也与其他细胞过程有关，包括转化，分化和凋亡。 PKR和RNA干扰（RNAI）途径之间也存在结构和功能连接。 PKR在潜在状态下合成，并在与dsRNA结合后被激活，以经历激活激酶的自磷酸化反应。反过来，活化的PKR磷酸化真核引发因子2a，从而抑制了病毒感染细胞中蛋白质合成的抑制。这种抗病毒途径的重要性是由病毒演变为对抗PKR的各种机制突出的。我们研究计划的广泛目标是定义分子激活和抑制PKR的机制。我们将定义使用定量的生物物理和结构方法来调节PKR活性的大分子复合物的形成的石化型，亲和力和自由能耦合。我们将确定来自流感病毒的乙型肝炎病毒和NS1的NS5A如何与PKR相互作用以逃避抗病毒药途径。这些影响毒力或赋予干扰素耐药性的蛋白质中的突变将与PKR结合和抑制作用相关。我们将确定肝素寡糖作为PKR激活剂的作用。合成寡糖的微阵列将被筛选，结构活动关系将是生成以定义PKR的新型小分子激活剂。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提供基础。