Viral Stress-inducible Gene Expression

病毒应激诱导基因表达

• 批准号: 6928886
• 负责人: GANES C. SEN
• 金额: $ 5.52万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-15 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6928886
• 关键词: JAK kinase; antisense nucleic acid; biological signal transduction; biotechnology; cell line; double stranded RNA; gene expression profiling; gene induction /repression; interferons; microarray technology; nuclear factor kappa beta; parainfluenza virus type 1; phosphatidylinositol 3 kinase; phosphorylation; protein kinase; protein protein interaction; small interfering RNA; toll like receptor; transcription factor; transfection; virus genetics; virus infection mechanism; virus replication

DESCRIPTION (provided by applicant): Viral infection of mammalian cells causes rapid and profound changes in the expression patterns of specific cellular genes. The proteins encoded by these genes, the viral stress-inducible genes (VSIG), are responsible for maintaining viral homeostasis in the infected cells. Many of the VSIG can also be induced in uninfected cells by interferons (IFN) or double-stranded (ds) RNA, common byproducts of viral infection. Here, we propose to identify the patterns of changes in cellular gene expression profiles in response to type I IFN, dsRNA and Sendai virus infection. Customized cDNA microarrays will be used to measure the levels of relevant cellular mRNAs in virus-infected or IFN/dsRNA treated cells and compared with the corresponding levels in untreated cells. Mutant cell lines, specifically defective in IFN or dsRNA-elicited signaling pathways, will be used to assess the contributions of these pathways in gene induction by Sendai virus. Furthermore, we propose to delineate the distinct signaling pathways used by the three agents to induce transcription of a common subset of VSIG that encode the P56 family of proteins. For this purpose, we will use mutant cell lines deficient in specific components of the Jak-STAT, the IRF3 and the NFkappaB signaling pathways as well as chemical inhibitors and siRNAs. DsRNA signaling uses Toll-like receptor 3 for inducing transcription of the P56 family of genes and it has recently been shown by us that receptor tyrosine phosphorylation is essential for this process. We propose to identify the roles of specific phosphotyrosine residues of TLR3 in mediating dsRNA-signaling that leads to the induction of P56 and other genes. The biochemical basis of the need for TLR3 phosphorylation will be investigated by examining activation of the relevant transcription factors and protein kinases in cells expressing Wt or mutant TLR3 proteins. These studies will illuminate the modes of cross-talk between the TLR and the Jak-STAT signaling pathways for appropriate gene induction in virus-infected cells.

描述（由申请人提供）：哺乳动物细胞的病毒感染会导致特定细胞基因的表达模式的快速而深刻的变化。 由这些基因编码的蛋白质，即病毒应激诱导基因（VSIG）负责维持感染细胞中的病毒稳态。 也可以通过干扰素（IFN）或双链（DS）RNA（病毒感染的常见副产物）在未感染的细胞中诱导许多VSIG。 在这里，我们提出，响应于I型IFN，DSRNA和仙台病毒感染而确定细胞基因表达谱的变化模式。 定制的cDNA微阵列将用于测量病毒感染或IFN/DSRNA处理的细胞中相关细胞mRNA的水平，并将其与未处理细胞中的相应水平进行比较。 突变细胞系，在IFN或DSRNA诱导的信号通路中特异性有缺陷，将用于评估仙台病毒基因诱导中这些途径的贡献。 此外，我们建议描述三种代理用于诱导p56蛋白质家族的VSIG的转录的不同信号通路。 为此，我们将使用突变细胞系在JAK-STAT，IRF3和NFKAPPAB信号通路以及化学抑制剂和siRNA的特定组件中缺乏突变细胞系。 DSRNA信号传导使用Toll样受体3来诱导p56基因家族的转录，并且最近已证明受体酪氨酸磷酸化对于此过程至关重要。 我们建议确定TLR3的特定磷酸酪氨酸残基在介导DSRNA信号中的作用，从而导致p56和其他基因的诱导。 将通过检查表达WT或突变体TLR3蛋白的细胞中相关转录因子和蛋白激酶的激活来研究对TLR3磷酸化需求的生化基础。 这些研究将阐明TLR和JAK-STAT信号通路之间的串扰模式，以便在病毒感染的细胞中适当的基因诱导。