Mechanism of Interferon Action

干扰素作用机制

• 批准号: 6983503
• 负责人: Charles E Samuel
• 金额: $ 27.44万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-05-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6983503
• 关键词: Adenoviridae; Poxviridae; adenosine deaminase; cell line; enzyme activity; enzyme induction /repression; enzyme mechanism; enzyme structure; enzyme substrate; gene expression; genetic promoter element; genetic transcription; host organism interaction; interferon alpha; interferon gamma; interferons; isozymes; laboratory mouse; laboratory rabbit; nucleic acid sequence; protein biosynthesis; protein localization; protein protein interaction; tissue /cell culture; virus RNA; virus replication

DESCRIPTION (provided by applicant): The Overall Objective of the proposed investigation is to elucidate the role of the interferon-inducible RNA-specific adenosine deaminase (ADAR1) in the actions that interferons (IFN) mediate on viral and host functions. The Specific Aims of our proposed continuation study of ADAR1 and double-stranded RNA editing in IFN action are as follows: (1) To further characterize the biochemical and biophysical properties of the IFN-inducible p150 ADAR1. To determine the mechanisms by which poxvirus E3L protein and adenovirus VA1 RNA antagonize ADAR1 enzymatic activity; to characterize the functional selectivity of dsRNA binding domains of ADAR1; and to determine the structural basis and functional significance of ADAR1 protein interactions and subcellular localizations in uninfected and virus-infected cells. (2) To further delineate the structure of the 5'-flanking regions of the ADAR1 gene required for IFN-inducible and basal transcriptional activity, including the Pi promoter responsible for inducible expression of transcripts encoding the p150 ADAR1 protein isoform and the Pb and Pc promoters responsible for constitutive expression of nuclear p110 ADAR1. To define the pathways leading to inducible transcriptional activation of ADAR1 expression in IFN-treated cells and in infected mice. (3) To further characterize the effect of singular stable expression of wild type and mutant forms of ADAR1 deaminase in cultured cells on virus multiplication including VA1 mutant adenovirus, E3L mutant vaccinia virus and recombinant measles virus, to generate a targeted deletion of the mouse ADAR1 inducible genomic locus, and to determine the effect of the ADAR1 p150 disruption on mouse phenotype and viral pathogenesis. The health relatedness of the proposed research stems from the likelihood that the work may contribute to a better understanding of regulatory mechanisms operative in normal cells as well as virus-infected cells. Furthermore, elucidation of the actions of IFN at the molecular level is of importance in view of potential applications of interferon in the clinic.

描述（由申请人提供）：拟议研究的总体目的是阐明干扰素诱导的RNA特异性腺苷脱氨酶（ADAR1）在干扰物（IFN）在病毒和宿主功能上介导的作用中的作用。在IFN作用中，我们提出的对ADAR1和双链RNA编辑的连续研究的具体目的如下：（1）进一步表征IFN诱导P150 ADAR1的生化和生物物理特性。为了确定痘病毒E3L蛋白和腺病毒VA1 RNA拮抗ADAR1酶活性的机制；表征adar1的dsRNA结合域的功能选择性；并确定ADAR1蛋白相互作用以及未感染和病毒感染细胞中亚细胞局部的结构基础和功能意义。 （2）进一步描绘了IFN诱导和基础转录活性所需的ADAR1基因的5'-频脉冲区域的结构，包括负责编码P150 ADAR1蛋白质相结合的转录物表达的PI启动子，以及负责核P110 Adar核P110 Adar的组成型表达的PB和PC启动子。定义导致IFN处理细胞和感染小鼠中ADAR1表达诱导转录激活的途径。 （3）进一步表征培养细胞中野生型和突变形式的奇异稳定表达在病毒繁殖中的影响的影响，包括VA1突变腺病毒，E3L突变体疫苗病毒和重新组合性病毒的影响，从和病毒发病机理。拟议研究的健康相关性源于该工作可能有助于更好地理解正常细胞和病毒感染细胞中的调节机制的可能性。此外，鉴于干扰素在诊所中的潜在应用，阐明IFN在分子水平上的作用至关重要。