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导致特定细胞基因的表达模式发生快速而深刻的变化。 这些基因编码的蛋白质，即病毒应激诱导基因（VSIG），负责维持受感染细胞中的病毒稳态。 许多 VSIG 也可以在未感染的细胞中由干扰素 (IFN) 或双链 (ds) RNA（病毒感染的常见副产物）诱导。 在这里，我们建议确定细胞基因表达谱响应 I 型 IFN、dsRNA 和仙台病毒感染的变化模式。 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，并与未处理细胞中的相应水平进行比较。 突变细胞系，尤其是 IFN 或 dsRNA 引发的信号通路缺陷的细胞系，将用于评估这些通路在仙台病毒基因诱导中的贡献。 此外，我们建议描述这三种药物用于诱导编码 P56 蛋白家族的 VSIG 常见子集转录的不同信号通路。 为此，我们将使用缺乏 Jak-STAT、IRF3 和 NFkappaB 信号通路特定成分以及化学抑制剂和 siRNA 的突变细胞系。 DsRNA 信号传导使用 Toll 样受体 3 来诱导 P56 基因家族的转录，我们最近表明受体酪氨酸磷酸化对于这一过程至关重要。 我们建议确定 TLR3 的特定磷酸酪氨酸残基在介导 dsRNA 信号传导中的作用，从而诱导 P56 和其他基因的诱导。 将通过检查表达 Wt 或突变 TLR3 蛋白的细胞中相关转录因子和蛋白激酶的激活来研究 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，以在病毒感染的细胞中进行适当的基因诱导。