The PDZ-binding motif of severe acute respiratory syndrome coronavirus envelope protein is a determinant of viral pathogenesis.
The PDZ-binding motif of severe acute respiratory syndrome coronavirus envelope protein is a determinant of viral pathogenesis.
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DOI：
10.1371/journal.ppat.1004320
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发表时间：
2014-08
期刊：
影响因子：
6.7
通讯作者：
Enjuanes L
中科院分区：
文献类型：
--
作者：
Jimenez-Guardeño JM;Nieto-Torres JL;DeDiego ML;Regla-Nava JA;Fernandez-Delgado R;Castaño-Rodriguez C;Enjuanes L
重组严重急性呼吸综合征冠状病毒(SARS-CoV)缺乏包膜(E)蛋白，在体内减毒。在这里，我们报告了E蛋白PDZ结合基序(PBM)，一个涉及蛋白质-蛋白质相互作用的结构域，是毒力的主要决定因素。通过使用反向遗传学消除SARS-CoV E蛋白PBM，可以减少感染亲本病毒时引发的免疫反应的有害加剧，并减少病毒的衰减。通过酵母双杂交、双向免疫共沉淀和共聚焦显微镜等三种互补策略，鉴定了在SARS冠状病毒感染过程中与E蛋白PBM结合的细胞蛋白Syntenin。在含有E蛋白PBM的病毒感染过程中，Syntenin从细胞核重新分布到细胞质中，激活p38MAPK，导致炎性细胞因子的过度表达。使用siRNAs沉默Syntenin导致感染SARS-CoV的细胞中p38MAPK活性降低，进一步加强了它们之间的功能关系。与亲本病毒相比，缺乏E蛋白pBM的SARS冠状病毒感染的小鼠肺内活性p38MAPK减少，导致炎性细胞因子的表达减少，病毒减毒。有趣的是，p38 MAPK抑制剂的应用导致感染SARS-CoV的小鼠存活率增加，证实了这一途径与SARS-CoV毒力的相关性。因此，E蛋白PBM是一个毒力区域，很可能是通过使用Syntenin作为p38MAPK诱导的炎症的介质来激活免疫病理的。SARS冠状病毒导致全球流行，感染8000人，死亡率约10%。对缺失E蛋白的重组SARS-CoV进行了体内减毒试验。E蛋白包含一个PDZ结合基序(PBM)，这个结构域可能参与与400多种细胞蛋白的相互作用，这突显了它在调节宿主细胞行为方面的相关性。为了分析该基序对毒力的贡献，构建了含有或不含有E蛋白pBM的重组病毒。与含有E蛋白PBM的病毒相比，缺乏E蛋白PBM的重组SARS冠状病毒对肺的损伤最小，且较弱，表明E蛋白PBM是一个毒力决定因素。蛋白质组学分析表明，E蛋白PBM在SARS冠状病毒感染过程中诱导有害的加重免疫反应，并与细胞蛋白Syntenin相互作用。有趣的是，在SARS冠状病毒感染过程中，Syntenin从细胞核重新分布到细胞质，激活p38MAPK，触发炎性细胞因子的过度表达。此外，使用siRNAs沉默Syntenin导致p38MAPK激活减少。此外，在感染SARS-CoV后，给予p38 MAPK抑制剂可以增加小鼠的存活率。这些结果表明，Syntenin和p38MAPK是减轻SARS冠状病毒感染时加剧的免疫反应的潜在治疗靶点。
A recombinant severe acute respiratory syndrome coronavirus (SARS-CoV) lacking the envelope (E) protein is attenuated in vivo. Here we report that E protein PDZ-binding motif (PBM), a domain involved in protein-protein interactions, is a major determinant of virulence. Elimination of SARS-CoV E protein PBM by using reverse genetics caused a reduction in the deleterious exacerbation of the immune response triggered during infection with the parental virus and virus attenuation. Cellular protein syntenin was identified to bind the E protein PBM during SARS-CoV infection by using three complementary strategies, yeast two-hybrid, reciprocal coimmunoprecipitation and confocal microscopy assays. Syntenin redistributed from the nucleus to the cell cytoplasm during infection with viruses containing the E protein PBM, activating p38 MAPK and leading to the overexpression of inflammatory cytokines. Silencing of syntenin using siRNAs led to a decrease in p38 MAPK activation in SARS-CoV infected cells, further reinforcing their functional relationship. Active p38 MAPK was reduced in lungs of mice infected with SARS-CoVs lacking E protein PBM as compared with the parental virus, leading to a decreased expression of inflammatory cytokines and to virus attenuation. Interestingly, administration of a p38 MAPK inhibitor led to an increase in mice survival after infection with SARS-CoV, confirming the relevance of this pathway in SARS-CoV virulence. Therefore, the E protein PBM is a virulence domain that activates immunopathology most likely by using syntenin as a mediator of p38 MAPK induced inflammation. SARS-CoV caused a worldwide epidemic infecting 8000 people with a mortality of about 10%. A recombinant SARS-CoV lacking the E protein was attenuated in vivo. The E protein contains a PDZ-binding motif (PBM), a domain potentially involved in the interaction with more than 400 cellular proteins, which highlights its relevance in modulating host-cell behavior. To analyze the contributions of this motif to virulence, recombinant viruses with or without E protein PBM were generated. Recombinant SARS-CoVs lacking E protein PBM caused minimal lung damage and were attenuated, in contrast to viruses containing this motif, indicating that E protein PBM is a virulence determinant. E protein PBM induces the deleterious exacerbated immune response triggered during SARS-CoV infection, and interacts with the cellular protein syntenin, as demonstrated using proteomic analyses. Interestingly, syntenin redistributed from nucleus to cytoplasm during SARS-CoV infection, activating p38 MAPK and triggering the overexpression of inflammatory cytokines. Furthermore, silencing of syntenin using siRNAs led to a decrease in p38 MAPK activation. In addition, administration of a p38 MAPK inhibitor led to an increase in mice survival after SARS-CoV infection. These results indicate that syntenin and p38 MAPK are potential therapeutic targets to reduce the exacerbated immune response during SARS-CoV infection.
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影响因子：
8
作者：
Boukerche, H.;Aissaoui, H.;Prevost, C.;Hirbec, H.;Das, S. K.;Su, Z-Z;Sarkar, D.;Fisher, P. B.
通讯作者：
Fisher, P. B.
影响因子：
3.7
作者：
Alvarez E;DeDiego ML;Nieto-Torres JL;Jiménez-Guardeño JM;Marcos-Villar L;Enjuanes L
通讯作者：
Enjuanes L
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y
DOI：
10.1056/nejmoa1306742
发表时间：
2013-08-01
期刊：
The New England journal of medicine
影响因子：
--
作者：
Assiri A;McGeer A;Perl TM;Price CS;Al Rabeeah AA;Cummings DA;Alabdullatif ZN;Assad M;Almulhim A;Makhdoom H;Madani H;Alhakeem R;Al-Tawfiq JA;Cotten M;Watson SJ;Kellam P;Zumla AI;Memish ZA;KSA MERS-CoV Investigation Team
通讯作者：
KSA MERS-CoV Investigation Team
影响因子：
14.8
作者：
Huang, Da Wei;Sherman, Brad T.;Lempicki, Richard A.
通讯作者：
Lempicki, Richard A.