A camel-derived MERS-CoV with a variant spike protein cleavage site and distinct fusion activation properties.
A camel-derived MERS-CoV with a variant spike protein cleavage site and distinct fusion activation properties.
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DOI：
10.1038/emi.2016.125
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发表时间：
2016-12-21
影响因子：
13.2
通讯作者：
Whittaker, Gary R.
中科院分区：
文献类型：
--
作者：
Millet, Jean Kaoru;Goldstein, Monty E.;Labitt, Rachael N.;Hsu, Hung-Lun;Daniel, Susan;Whittaker, Gary R.
中东呼吸综合征冠状病毒（MERS-CoV）继续在人类和骆驼中传播，该病毒的起源和进化尚不清楚。在这里，我们对2013年中东呼吸综合征爆发早期发现的骆驼源性MERS- cov （NRCE-HKU205）的刺突蛋白进行了表征。NRCE-HKU205刺突蛋白在S2 ‘融合激活位点上有一个变异的切割基序——值得注意的是，在关键的P1 ’切割位点位置上，异亮氨酸取代了其他不变的丝氨酸。正如荧光肽裂解和western blot检测所显示的那样，取代导致了furin介导的裂解缺失。细胞-细胞融合和假型病毒的感染性实验表明，S2 '的替换减少了刺突介导的融合和病毒的进入。然而，组织蛋白酶和胰蛋白酶样蛋白酶的激活仍然存在，尽管与EMC/2012原型人菌株相比效率大大降低。我们发现NRCE-HKU205具有更有限的融合激活特性，可能导致更有限的病毒趋向性，并且可能代表MERS-CoV生态和进化复杂模式的中间物。
Middle East respiratory syndrome coronavirus (MERS-CoV) continues to circulate in both humans and camels, and the origin and evolution of the virus remain unclear. Here we characterize the spike protein of a camel-derived MERS-CoV (NRCE-HKU205) identified in 2013, early in the MERS outbreak. NRCE-HKU205 spike protein has a variant cleavage motif with regard to the S2′ fusion activation site—notably, a novel substitution of isoleucine for the otherwise invariant serine at the critical P1′ cleavage site position. The substitutions resulted in a loss of furin-mediated cleavage, as shown by fluorogenic peptide cleavage and western blot assays. Cell–cell fusion and pseudotyped virus infectivity assays demonstrated that the S2′ substitutions decreased spike-mediated fusion and viral entry. However, cathepsin and trypsin-like protease activation were retained, albeit with much reduced efficiency compared with the prototypical EMC/2012 human strain. We show that NRCE-HKU205 has more limited fusion activation properties possibly resulting in more restricted viral tropism and may represent an intermediate in the complex pattern of MERS-CoV ecology and evolution.
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DOI：
10.1038/nrmicro.2016.81
发表时间：
2016-08
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
de Wit E;van Doremalen N;Falzarano D;Munster VJ
通讯作者：
Munster VJ
通过混合方法计算预测弗林蛋白酶切割位点并了解疾病的潜在机制
影响因子：
4.6
作者：
Tian, Sun;Wang Huajun;Wu, Jianhua
通讯作者：
Wu, Jianhua
影响因子：
14.9
作者：
Rawlings ND;Barrett AJ;Finn R
通讯作者：
Finn R
影响因子：
56.9
作者：
Sabir, Jamal S. M.;Lam, Tommy T. -Y.;Guan, Yi
通讯作者：
Guan, Yi
DOI：
10.1073/pnas.1407087111
发表时间：
2014-10-21
影响因子：
11.1
作者：
Millet, Jean Kaoru;Whittaker, Gary R.
通讯作者：
Whittaker, Gary R.