Generating reagents to explore the anitviral potential of TRIM family proteins

描述（由申请人提供）：TRIM蛋白家族介导了许多最近报道的抗多种病毒的抗病毒活性。对这些活性的最佳研究是TRIM 51蛋白在感染的早期阶段通过结合存在于病毒衣壳核心上的决定簇来抑制某些逆转录病毒（包括HIV-1）复制的能力。最近的数据表明，其他TRIM家族蛋白可以在病毒复制周期的其他步骤中影响HIV-1复制。许多研究表明，TRIM家族蛋白也可以负调节多种病毒的复制，包括腺病毒、疱疹病毒和流感病毒，尽管这些活性仍然缺乏表征。最近发现许多TRIM家族蛋白的表达响应于干扰素治疗而上调，这也表明TRIM家族蛋白代表先天免疫系统的细胞内元件，其作用是抑制细胞内病原体的复制。由于TRIM 51在细胞中倾向于多聚成高级结构，因此这种蛋白质的生化分析异常困难。其他TRIM家族成员可能也是如此。我以前的研究利用定量荧光显微镜观察和量化TRIM 51和HIV-1病毒复合物在细胞质中发生的相互作用。该项目将开发诱导表位标记的TRIM家族成员表达的逆转录病毒载体，并使用这些载体产生稳定表达这些蛋白质的细胞系。这些载体和细胞系将可用于鉴定具有抗多种病毒的抗病毒活性的TRIM蛋白，并揭示细胞中病毒和TRIM家族蛋白之间发生的新型相互作用。一旦鉴定，这些相同的载体和细胞系将允许可视化TRIM蛋白和受TRIM活性影响的病毒蛋白和/或遗传元件之间发生的相互作用。这将促进我的实验室研究的新领域，并增加我们对TRIM家族蛋白质抗病毒活性的理解。 TRIM家族蛋白的抗病毒潜力是先天免疫系统研究的一个新兴领域。这项研究将开发试剂来鉴定可以抑制多种病毒复制的TRIM蛋白。这些试剂还将允许使用定量荧光显微镜鉴定和表征细胞中TRIM蛋白和病毒决定簇之间发生的相互作用。

DESCRIPTION (provided by applicant):The TRIM family of proteins mediates a number of recently reported antiviral activities mediated against diverse viruses. The best studied of these activities is the ability of the TRIM51 protein to inhibit the replication of certain retroviruses, including HIV-1, by binding determinants present on the viral capsid core during early stages of infection. Recent data suggests that other TRIM family proteins can affect HIV-1 replication at other steps in the viral replication cycle. Numerous studies suggest that TRIM family proteins can also negatively regulate the replication of a diverse range of viruses, including adenoviruses, herpesviruses and influenza viruses, although these activities remain poorly characterized. The recent finding that the expression of many TRIM family proteins are upregulated in response to interferon treatment also indicates that the TRIM family proteins represent an intracellular element of the innate immune system, acting to inhibit the replication of intracellular pathogens. Because of the tendency of TRIM51 to multimerize into high-order structures in cells, biochemical analysis of this protein has been unusually difficult. This will likely be true of other TRIM family members. My previous studies have utilized quantitative fluorescent microscopy to visualize and quantify the interactions occurring between TRIM51 and HIV-1 viral complexes in the cytoplasm. This project would develop retroviral vectors that induce the expression of epitope tagged TRIM family members and use these vectors to generate cell lines stably expressing these proteins. These vectors and cell lines will be useful in identifying TRIM proteins with antiviral activity against diverse viruses and expose novel interactions occurring in cells between viruses and TRIM family proteins. Once identified, these same vectors and cell lines will allow for the visualization of interactions occurring between TRIM proteins and viral proteins and/or genetic elements affected by TRIM activity. This will catalyze new areas of research for my lab and increase our understanding of the antiviral activity of TRIM family proteins. PUBLIC HEALTH RELEVANCE: The antiviral potential of TRIM family proteins is an emerging area of innate immune system research. This research will develop reagents to identify TRIM proteins that can inhibit the replication of diverse viruses. These reagents will also allow the interactions occurring between TRIM proteins and viral determinants in cells to be identified and characterized using quantitative, fluorescent microscopy.