Delineating differential cellular stress signaling responses induced by reovirus

描述呼肠孤病毒诱导的差异细胞应激信号反应

• 批准号: RGPIN-2018-05003
• 负责人: Coombs, Kevin
• 金额: $ 2.62万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=713475
• 关键词: Delineating; differential; cellular; stress; signaling

Mammalian cells are complex entities consisting of tens of thousands of genes. These genes are expressed at different times and to different extents to produce the hundreds of thousands to millions of different cell proteins that coordinate all cellular activities involved in development, homeostasis, repair and immunity. This coordination is regulated by cell signaling pathways. Viruses use and disrupt cell signaling pathways. Viruses also are much simpler than cells, being composed of only a few genes and proteins. Thus, viruses continue to serve as superb models for understanding fundamental aspects of cell biology, including signaling pathways. My long-term goals are to delineate these signaling processes; to determine how a non-pathogenic mammalian reovirus (MRV) affects cellular signaling, how cell signaling perturbations enhance or disrupt viral processes, to determine which viral genes induce these signaling responses, to determine if, and which, post-translational modifications promote or abrogate signaling, and to precisely elucidate what molecular regions and domains of specific viral proteins are responsible for these signaling processes. These studies will improve our understanding of basic cell biology and of a critically important master regulator of cellular processes. My lab group previously published mass spectrometry-based analyses of MRV-infected cells and showed that the two most common MRV serotypes (type 1 Lang, T1L and type 3 Dearing, T3D) induced differential cellular protein responses. We also have preliminary quantitative aptamer-based results showing similar differential responses in cell signaling pathways. My group will continue its programmatic study of how MRV affect cellular signaling. Our short-term goals are to confirm and extend the preliminary aptamer studies to identify cell proteins that are significantly dysregulated, screen these significant proteins by RNA interference to knock down the corresponding genes to determine their effects upon virus and cellular functions, and exploit a novel genetic characteristic of MRV to map which regions of which viral proteins induce these signaling perturbations. Collectively, these studies will allow us to delineate important molecular components of fundamental cellular processes. The HQP who are trained in my laboratory have, and will continue, to benefit from our expertise in molecular biology and proteomics, and will acquire in-demand skill sets that will be broadly applicable in a wide variety of biologically important systems. These new HQP will be well equipped for careers in basic scientific research focused on biochemistry, biotechnology, genetics, molecular biology, and proteomics.

哺乳动物细胞是由成千上万个基因组成的复杂实体。这些基因在不同的时间和不同的程度上表达，以产生数十万到数百万不同的细胞蛋白，这些蛋白协调涉及发育、内稳态、修复和免疫的所有细胞活动。这种协调受细胞信号通路的调节。病毒利用并破坏细胞信号通路。病毒也比细胞简单得多，只由几个基因和蛋白质组成。因此，病毒仍然是理解细胞生物学的基本方面，包括信号通路的极好模型。 我的长期目标是描绘这些信号传递过程；确定非致病性哺乳动物呼肠孤病毒(MRV)如何影响细胞信号传递；细胞信号干扰如何增强或干扰病毒过程；确定哪些病毒基因诱导这些信号反应；确定翻译后修饰是否以及哪些翻译后修饰促进或取消信号传递；以及准确地阐明特定病毒蛋白的哪些分子区域和结构域负责这些信号传递过程。 这些研究将提高我们对基本细胞生物学的理解，以及对细胞过程中一个至关重要的主要调节因子的理解。 我的实验室小组之前发表了基于质谱学的MRV感染细胞分析，并表明两种最常见的MRV血清型(1型Lang，T1L和3型Dearing，T3d)诱导不同的细胞蛋白反应。我们也有基于适体的初步定量结果，显示细胞信号通路中类似的差异反应。 我的团队将继续对MRV如何影响细胞信号进行程序性研究。我们的短期目标是确认和扩大初步的适配子研究，以确定显著失调的细胞蛋白，通过RNA干扰筛选这些重要蛋白来敲除相应的基因，以确定它们对病毒和细胞功能的影响，并利用MRV的一个新的遗传特征来定位哪些病毒蛋白区域引起这些信号干扰。总而言之，这些研究将使我们能够描绘基本细胞过程的重要分子成分。在我的实验室接受培训的HQP已经并将继续受益于我们在分子生物学和蛋白质组学方面的专业知识，并将获得广泛适用于各种生物重要系统的需求技能。这些新的HQP将为专注于生物化学、生物技术、遗传学、分子生物学和蛋白质组学的基础科学研究的职业生涯做好准备。