Antiviral Activity of Guanylate-Binding Proteins and Viral Countermeasures

鸟苷酸结合蛋白的抗病毒活性和病毒对策

• 批准号: 400912104
• 负责人: Professor Dr. Frank Kirchhoff
• 金额: --
• 依托单位: Institut für Molekulare Virologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/400912104?language=en
• 关键词: Antiviral; Activity; Guanylate; Binding; Proteins

Guanylate-binding proteins (GBPs) are interferon-inducible GTPases known to play key roles in protective innate immunity against bacteria and protozoa. Humans encode seven GBP paralogs. Using evolutionary genomics and protein interaction data, we found that one of them (GBP5) shares characteristics of known antiretroviral restriction factors. Functional analyses confirmed that GBP5 restricts HIV-1 and other retroviruses by interfering with processing and virion incorporation of the viral envelope (Env) glycoprotein. The inhibitory activity of GBP5 requires isoprenylation-dependent Golgi-localization but not its GTPase function. GBP5 efficiently restricts HIV-1 in human macrophages, and some brain-derived HIV-1 strains evade this antiviral effect by an unusual tradeoff mechanism: mutations in the vpu initiation codon increase Env expression at the cost of Vpu function. Our ongoing studies demonstrate that GBP5 is also inducible by L-27 and TCR-CD3 signaling, that GBP2 also restricts lentiviral replication, and that the antiviral activity of GBP2 and GBP5 is evolutionarily conserved. In the proposed project, we want to define the exact mechanism(s) underlying restriction as well as lentiviral evasion or counteraction. Initially, we will map the amino acid residues that are critical for the antiviral activity of GBP5 and analyze their effect on Env processing, trafficking and virion incorporation. Furthermore, we will determine how the susceptibility of HIV-1 and HIV-2 to GBP2/5 inhibition evolved after transmission of SIVs from great apes and sooty mangabeys to elucidate whether GBPs constitute a barrier to successful zoonotic viral transmission. These studies will also reveal how these viruses evade or counteract the inhibitory effect of GBP2/5. Another focus will be the targeted induction of GBP2/5 expression in in primary viral target cells by different cytokines. Finally, our preliminary results show that GBP2 and GBP5 do not only inhibit retroviruses. Thus, we will analyse the effects of GBP proteins on a variety of viral pathogens, such as Influenza, Ebola, Marburg, Lyssa, Respiratory Syncytial and SARS Coronavirus. The results will yield important insights into antiviral defence mechanisms and viral countermeasures.

鸟苷酸结合蛋白 (GBP) 是干扰素诱导型 GTP 酶，已知在针对细菌和原生动物的保护性先天免疫中发挥关键作用。人类编码七种英镑旁系同源物。利用进化基因组学和蛋白质相互作用数据，我们发现其中之一（GBP5）具有已知抗逆转录病毒限制因子的特征。功能分析证实，GBP5 通过干扰病毒包膜 (Env) 糖蛋白的加工和病毒体掺入来限制 HIV-1 和其他逆转录病毒。 GBP5 的抑制活性需要异戊二烯化依赖性高尔基体定位，但不需要其 GTP 酶功能。 GBP5 有效地限制人类巨噬细胞中的 HIV-1，一些脑源性 HIV-1 菌株通过一种不寻常的权衡机制来逃避这种抗病毒作用：vpu 起始密码子的突变以牺牲 Vpu 功能为代价增加 Env 表达。我们正在进行的研究表明，GBP5 也可由 L-27 和 TCR-CD3 信号传导诱导，GBP2 也限制慢病毒复制，并且 GBP2 和 GBP5 的抗病毒活性在进化上是保守的。在拟议的项目中，我们希望定义限制以及慢病毒逃避或对抗的确切机制。首先，我们将绘制对 GBP5 抗病毒活性至关重要的氨基酸残基，并分析它们对 Env 加工、运输和病毒颗粒掺入的影响。此外，我们将确定在类人猿和乌白眉猴传播 SIV 后，HIV-1 和 HIV-2 对 GBP2/5 抑制的敏感性如何演变，以阐明 GBP 是否构成人畜共患病毒成功传播的障碍。这些研究还将揭示这些病毒如何逃避或抵消 GBP2/5 的抑制作用。另一个焦点是不同细胞因子在原代病毒靶细胞中定向诱导 GBP2/5 表达。最后，我们的初步结果表明 GBP2 和 GBP5 不仅抑制逆转录病毒。因此，我们将分析GBP蛋白对多种病毒病原体的影响，例如流感、埃博拉、马尔堡、狂犬病、呼吸道合胞病毒和SARS冠状病毒。研究结果将为抗病毒防御机制和病毒对策提供重要见解。

项目成果

Structural basis for GTP-induced dimerization and antiviral function of guanylate-binding proteins

GTP 诱导的鸟苷酸结合蛋白二聚化和抗病毒功能的结构基础

• DOI: 10.1073/pnas.2022269118
• 发表时间: 2021-04-13
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Cui，Wen;Braun，Elisabeth;Yang，Haitao
• 通讯作者: Yang，Haitao