Aminopeptidase N is not required for porcine epidemic diarrhea virus cell entry.
Aminopeptidase N is not required for porcine epidemic diarrhea virus cell entry.
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DOI：
10.1016/j.virusres.2017.03.018
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发表时间：
2017-05-02
期刊：
影响因子：
5
通讯作者：
Bosch BJ
中科院分区：
文献类型：
--
作者：
Li W;Luo R;He Q;van Kuppeveld FJM;Rottier PJM;Bosch BJ
猪 APN 在细胞中的过度表达不会导致对 PEDV 的易感性。 PEDV 易感细胞中的 APN 表达敲除对 PEDV 感染没有影响。结果表明 APN 对于 PEDV 进入细胞不是必需的。猪流行性腹泻病毒（PEDV）是一种新出现的致病性冠状病毒，给养猪业造成重大经济负担。该病毒感染肠上皮并引起绒毛萎缩，导致腹泻和脱水。病毒刺突 (S) 表面糖蛋白通过其 S1 亚基与宿主细胞受体的相互作用是感染的第一步，也是病毒趋向性的主要决定因素。至于其他几种α冠状病毒，包括猪传染性胃肠炎病毒（TGEV）和人类冠状病毒229E（HCoV-229E），据报道氨肽酶N（APN）蛋白是PEDV的功能受体。在这项研究中，我们研究了 APN 作为受体的作用。我们发现猪 APN 的过度表达使 MDCK 细胞对 TGEV 敏感，但对 PEDV 不敏感。与 TGEV-S1 不同，PEDV-S1 始终不表现出与细胞表面表达的 APN 或 APN 可溶性形式的结合。此外，将这些病毒与可溶性APN预孵育或用可溶性TGEV-S1预处理表达APN的ST细胞可阻断TGEV感染，但对PEDV感染没有影响。综合观察结果表明，PEDV 感染不需要 APN。为了明确证明这一结论，我们应用 CRISPR/Cas9 基因组工程敲除 PEDV 易感猪 (ST) 和人类细胞系（Huh7 和 HeLa）中的 APN 表达。因此，这些细胞不再在其表面结合 TGEV-S1 和 HCoV-229E-S1，并且能够抵抗相应病毒的感染。然而，APN 表达的基因消除对 PEDV 的感染性没有影响，这表明 APN 对于 PEDV 进入细胞并不是必需的。
Overexpression of porcine APN in cells does not confer susceptibility to PEDV. Knockout APN expression in PEDV-susceptible cells has no effect on PEDV infection. Results demonstrate that APN is not essential for PEDV cell entry. Porcine epidemic diarrhea virus (PEDV) is an emerging pathogenic coronavirus that causes a significant economic burden to the swine industry. The virus infects the intestinal epithelium and causes villous atrophy, resulting in diarrhea and dehydration. Interaction of the viral spike (S) surface glycoprotein − through its S1 subunit − with the host cell receptor is the first step in infection and the main determinant for virus tropism. As for several other alphacoronaviruses including the porcine transmissible gastroenteritis virus (TGEV) and the human coronavirus 229E (HCoV-229E), the aminopeptidase N (APN) protein was reported to be a functional receptor for PEDV. In this study we examined the role of APN as a receptor. We show that overexpression of porcine APN renders MDCK cells susceptible to TGEV, but not to PEDV. Consistently, unlike TGEV-S1, PEDV-S1 exhibited no binding to cell-surface expressed APN or to a soluble version of APN. Moreover, preincubation of these viruses with soluble APN or pretreatment of APN expressing ST cells with soluble TGEV-S1 blocked TGEV infection, but had no effect on infection by PEDV. The combined observations indicated that APN is not required for PEDV infection. To definitively prove this conclusion, we applied CRISPR/Cas9 genome engineering to knock out APN expression in PEDV-susceptible porcine (ST) and human cell lines (Huh7 and HeLa). As a consequence these cells no longer bound TGEV-S1 and HCoV-229E-S1 at their surface and were resistant to infection by the corresponding viruses. However, genetic ablation of APN expression had no effect on their infectability by PEDV, demonstrating that APN is not essential for PEDV cell entry.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
3.7
作者：
Li C;Li Z;Zou Y;Wicht O;van Kuppeveld FJ;Rottier PJ;Bosch BJ
通讯作者：
Bosch BJ
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
猪氨肽酶 N 是 PEDV 冠状病毒的功能受体
影响因子：
3.7
作者：
Li, B. X.;Ge, J. W.;Li, Y. J.
通讯作者：
Li, Y. J.
DOI：
10.1073/pnas.1210123109
发表时间：
2012-10-30
影响因子：
11.1
作者：
Chen, Lang;Lin, Yi-Lun;Li, Fang
通讯作者：
Li, Fang