Impact of viral and cellular protein phosphorylation on the regulation of innate antiviral immune responses against influenza A virus infections

病毒和细胞蛋白磷酸化对甲型流感病毒感染先天抗病毒免疫反应调节的影响

• 批准号: 316675205
• 负责人: Professor Dr. Stephan Ludwig
• 金额: --
• 依托单位: Abteilung für Molekulare Virologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/316675205?language=en
• 关键词: Impact; viral; cellular; protein; phosphorylation

Influenza viruses are small RNA viruses with a limited coding capacity. Therefore, viral proteins have to fulfill multiple functions in the infected cell. This implies that there must be mechanisms allowing for the control of multi-functionality in a spatio-temporal manner. We hypothesized that dynamic phosphorylation of viral and cellular proteins is a so far underestimated layer of regulation of virus propagation and innate immune responses to infection. To address this, we followed a strategy to focus for in depth functional analysis on a set of individual functionally related viral and cellular phosphoproteins identified by our own phospho-proteomic screen with a suspected function in the innate immune response and viral replication. In the previous funding period, we were not only able to describe the regulatory function of phosphorylation of a range of viral and cellular proteins, but also identified targets that may be used for novel host-directed anti-infective strategies. We have now performed additional phospho-proteome and kinome screenings, which identified further novel phospho-sites of the viral IFN-antagonistic protein NS1 as well as kinases and kinase regulators that are central in a virusrelated functional network analysis. We now want to build on these findings and enhance our studies by functional characterization of the three newly identified sites in NS1, this time also considering dynamic interaction with neighboring phospho-acceptor sites. Furthermore, we will unravel the role of two particular kinases, Cdk1 and PKA, that are differentially phosphorylated and activated during the virus replication cycle and also are linked to innate immune responses and described to phosphorylate NS1. This study will provide deeper knowledge about the regulatory host-virus interplay and is key to uncover new leads for pharmaceutical intervention.

流感病毒是编码能力有限的小RNA病毒。因此，病毒蛋白必须在感染细胞中完成多种功能。这意味着必须有一种机制允许以时空方式控制多功能。我们假设病毒和细胞蛋白的动态磷酸化是迄今为止被低估的病毒繁殖和对感染的先天免疫反应的调节层。为了解决这个问题，我们采用了一种策略，对我们自己的磷酸化蛋白质组学筛选确定的一组与先天免疫反应和病毒复制有关的单个功能相关的病毒和细胞磷酸化蛋白进行了深入的功能分析。在之前的资助期内，我们不仅能够描述一系列病毒和细胞蛋白磷酸化的调控功能，而且还确定了可能用于新型宿主定向抗感染策略的靶标。我们现在进行了额外的磷酸化蛋白质组和激酶组筛选，进一步确定了病毒ifn拮抗蛋白NS1的新的磷酸化位点，以及在病毒相关功能网络分析中处于中心位置的激酶和激酶调节因子。我们现在希望在这些发现的基础上，通过对NS1中三个新发现位点的功能表征来加强我们的研究，这次我们还考虑了与邻近磷酸化受体位点的动态相互作用。此外，我们将揭示两种特定激酶Cdk1和PKA的作用，它们在病毒复制周期中被不同地磷酸化和激活，也与先天免疫反应有关，并被描述为磷酸化NS1。这项研究将提供关于宿主-病毒相互作用的更深入的知识，并且是发现药物干预新线索的关键。