Irreversible inactivation of ISG15 by a viral leader protease enables alternative infection detection strategies.
Irreversible inactivation of ISG15 by a viral leader protease enables alternative infection detection strategies.
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DOI：
10.1073/pnas.1710617115
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发表时间：
2018-03-06
影响因子：
11.1
通讯作者：
Komander D
中科院分区：
文献类型：
--
作者：
Swatek KN;Aumayr M;Pruneda JN;Visser LJ;Berryman S;Kueck AF;Geurink PP;Ovaa H;van Kuppeveld FJM;Tuthill TJ;Skern T;Komander D
了解病毒逃避宿主免疫的机制对开发抗病毒药物和病毒检测策略至关重要。泛素和泛素样修饰在细胞先天免疫和感染反应中是至关重要的，并且经常被病毒蛋白抑制。我们在这里发现了一种以前未知的病毒逃避机制。一种名为Lbpro的病毒蛋白酶可以不完全地从蛋白质中去除泛素和泛素样蛋白ISG15。虽然这一策略有效且不可逆转地关闭了这些修饰系统，但它使为病毒检测中的泛素研究开发的工具和技术得以重新利用。具体地说，我们发现口蹄疫病毒感染可以使用为泛素质谱研究开发的抗glygly抗体检测。在对病毒感染的反应中，细胞产生一种依赖于ISG15和泛素翻译后修饰的强效炎症反应。许多病毒使用去泛素酶和去isgyylase来逆转这些修饰并对抗宿主信号传导过程。我们在这里揭示了口蹄疫病毒（FMDV）的先导蛋白酶Lbpro以前所未有的方式靶向ISG15，并在较小程度上靶向泛素。与典型的去isgyylases不同的是，它水解GlyGly基序c末端后的异肽链，而Lbpro则是在GlyGly基序之前切割肽键。因此，GlyGly二肽仍然附着在底物Lys上，并且被切割的ISG15无法再偶联。Lbpro结合到工程ISG15自杀探针的晶体结构揭示了ISG15蛋白水解的分子基础。重要的是，针对泛素蛋白组学开发的抗glygly抗体能够检测病毒感染期间的Lbpro切割产物。这为基于宿主衍生的不可变表位检测口蹄疫病毒感染开辟了道路。
An understanding of the mechanisms by which viruses evade host immunity is essential to the development of antiviral drugs and viral detection strategies. Ubiquitin and ubiquitin-like modifications are crucial in cellular innate immune and infection responses and are often suppressed by viral proteins. We here identify a previously unknown mechanism of viral evasion. A viral protease, Lbpro, removes ubiquitin and the ubiquitin-like protein ISG15 incompletely from proteins. While this strategy efficiently and irreversibly shuts down these modification systems, it enables repurposing of tools and technologies developed for ubiquitin research in virus detection. Specifically, we show that foot-and-mouth disease virus infection can be detected using an anti-GlyGly antibody developed for ubiquitin mass spectrometry research. In response to viral infection, cells mount a potent inflammatory response that relies on ISG15 and ubiquitin posttranslational modifications. Many viruses use deubiquitinases and deISGylases that reverse these modifications and antagonize host signaling processes. We here reveal that the leader protease, Lbpro, from foot-and-mouth disease virus (FMDV) targets ISG15 and to a lesser extent, ubiquitin in an unprecedented manner. Unlike canonical deISGylases that hydrolyze the isopeptide linkage after the C-terminal GlyGly motif, Lbpro cleaves the peptide bond preceding the GlyGly motif. Consequently, the GlyGly dipeptide remains attached to the substrate Lys, and cleaved ISG15 is rendered incompetent for reconjugation. A crystal structure of Lbpro bound to an engineered ISG15 suicide probe revealed the molecular basis for ISG15 proteolysis. Importantly, anti-GlyGly antibodies, developed for ubiquitin proteomics, are able to detect Lbpro cleavage products during viral infection. This opens avenues for infection detection of FMDV based on an immutable, host-derived epitope.
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DOI：
10.1126/science.1227026
发表时间：
2012-11-23
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Choy A;Dancourt J;Mugo B;O'Connor TJ;Isberg RR;Melia TJ;Roy CR
通讯作者：
Roy CR
DOI：
10.1002/pro.2807
发表时间：
2015-12
期刊：
Protein science : a publication of the Protein Society
影响因子：
--
作者：
Aumayr M;Fedosyuk S;Ruzicska K;Sousa-Blin C;Kontaxis G;Skern T
通讯作者：
Skern T
DOI：
10.1126/science.1224026
发表时间：
2012-09-28
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bogunovic D;Byun M;Durfee LA;Abhyankar A;Sanal O;Mansouri D;Salem S;Radovanovic I;Grant AV;Adimi P;Mansouri N;Okada S;Bryant VL;Kong XF;Kreins A;Velez MM;Boisson B;Khalilzadeh S;Ozcelik U;Darazam IA;Schoggins JW;Rice CM;Al-Muhsen S;Behr M;Vogt G;Puel A;Bustamante J;Gros P;Huibregtse JM;Abel L;Boisson-Dupuis S;Casanova JL
通讯作者：
Casanova JL
DOI：
10.1073/pnas.1013388108
发表时间：
2011-02-08
影响因子：
11.1
作者：
James, Terrence W.;Frias-Staheli, Natalia;Mark, Brian L.
通讯作者：
Mark, Brian L.
影响因子：
30.3
作者：
Isaacson MK;Ploegh HL
通讯作者：
Ploegh HL