Cellular determinants of apoptosis in virus-infected cells

病毒感染细胞凋亡的细胞决定因素

基本信息

批准号：
8690329
负责人：
GANES C. SEN
金额：
$ 17.9万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8690329
关键词：
1-Phosphatidylinositol 3-Kinase Ablation Adenoviruses Antiviral Agents Antiviral Response Apoptosis Apoptotic BIRC4 gene Binding Biochemical Biochemical Genetics Cells Cessation of life Cytoplasmic Receptors DNA Viruses Defense Mechanisms Epidermal Growth Factor Receptor Family Funding Genes Genetic Genetic Transcription Goals Health Human Immune response Infection Interferon Type I Interferons Investigation Knock-in Mouse Knock-out Ligase Lysine Maintenance Mammals Maps Measures Mediating Mitochondria Modification Mouse Strains Mus Mutant Strains Mice Names Nature Organism Outcome Pathogenesis Pathway interactions Permeability Phosphorylation Physiological Play Process Proteins RNA Viruses Regulation Research Risk Role Serine Signal Pathway Signal Transduction Societies System Testing Toll-like receptors Transcriptional Activation Ubiquitination Viral Viral Pathogenesis Virus Virus Diseases Virus Replication X-linked IAP activating transcription factor arm base cell type combat genetically modified cells helicase in vivo member mutant novel pro-apoptotic protein promoter public health relevance response suicidal transcription factor viral DNA

项目摘要

DESCRIPTION (provided by applicant): The goal of the project is to investigate how a newly discovered antiviral pathway is activated and regulated and what its role is in determining viral pathogenesis and persistence. The type I interferon (IFN) system plays a major role in innate antiviral response. Virus infection activates the transcription factor IRF-3, which is responsible for the induction of IFN and other antiviral proteins. Recent investigation by us has revealed that IRF-3 also activates an IFN-independent pro-apoptotic pathway, named RIPA (RIG-I-activated IRF-3-mediated Pathway of Apoptosis). To trigger RIPA, activated IRF-3 binds Bax and translocates it to mitochondria to cause apoptosis; the two functions of IRF-3 are genetically separable. RIPA is activated by many RNA and DNA viruses and inhibits both viral replication and pathogenesis. RIPA is temporally regulated through the action of XIAP and PI3 kinase, which is also activated upon virus infection. In the absence of RIPA, viruses establish persistent infection. Here we propose to investigate RIPA further. In Aim 1, we will investigate how IRF-3 is activated in RIPA as a consequence of phosphorylation of specific serine residues and ubiquitynation of specific lysine residues. Genetic and biochemical analyses will be used for this purpose. In Aim 2, we will investigate how viruses evade RIPA temporarily, by triggering negative regulation of RIPA though activation of the epidermal growth factor receptor, which in turn activates PI3 kinase. In Aim 3, we will investigate the physiological relevance of RIPA in determining the outcome of virus infection. Mutant cells defective in one, but not the other, action of IRF-3, will be used for measuring the role of RIPA on the efficiency of virus replication and establishing viral persistence. Finally, genetically modified mice, including new IRF-3 mutant knock-in mice which have been generated for this project, will be used to assess the in vivo contribution of RIPA in controlling viral pathogenesis.

描述（由申请人提供）：该项目的目的是研究新发现的抗病毒途径是如何激活和调节的，以及其在确定病毒发病机理和持久性方面的作用。 I型干扰素（IFN）系统在先天抗病毒反应中起着重要作用。病毒感染激活了转录因子IRF-3，该因子是导致IFN和其他抗病毒药蛋白诱导的。我们最近的调查显示 IRF-3还激活了IFN独立的促凋亡途径，称为RIPA（RIG-I-Actived IRF-3介导的凋亡途径）。为了触发RIPA，激活的IRF-3结合了Bax并将其易位到线粒体引起细胞凋亡。 IRF-3的两个功能在遗传上是可分离的。 RIPA被许多RNA和DNA病毒激活，并抑制病毒复制和发病机理。 RIPA通过XIAP和PI3激酶的作用在时间上调节，这也被病毒感染激活。在缺乏RIPA的情况下，病毒建立了持续的感染。在这里，我们建议进一步研究RIPA。在AIM 1中，我们将研究如何在RIPA中激活IRF-3，这是由于特定丝氨酸残基的磷酸化和特定赖氨酸残基的泛素化的结果。遗传和生化分析将用于此目的。在AIM 2中，我们将通过激活表皮生长因子受体的激活来触发RIPA的阴性调节，研究病毒如何暂时逃避RIPA，这又激活了PI3激酶。在AIM 3中，我们将研究RIPA在确定病毒感染结果方面的生理相关性。突变细胞在一个但不存在IRF-3的作用中有缺陷，将用于测量RIPA对病毒复制效率的作用并建立病毒持久性。最后，将使用该项目生成的新的IRF-3突变型小鼠（包括新的IRF-3突变型小鼠）来评估RIPA在控制病毒发病机理中的体内贡献。