ATP1A1-Mediated Src Signaling Inhibits Coronavirus Entry into Host Cells
ATP1A1-Mediated Src Signaling Inhibits Coronavirus Entry into Host Cells
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DOI：
10.1128/jvi.03274-14
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发表时间：
2015-04-01
影响因子：
5.4
通讯作者：
de Haan, Cornelis A. M.
中科院分区：
文献类型：
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作者：
Burkard, Christine;Verheije, Monique H.;de Haan, Cornelis A. M.
除了转运离子外，多亚基Na+，K+-ATP酶还通过将强心类固醇（CTS）结合诱导的信号传递到细胞中来发挥功能。在这项研究中，我们分析了Na+，K+-ATP酶，特别是其ATP 1A 1 α亚基在冠状病毒（CoV）感染过程中的作用。作为对照，包括水泡性口炎病毒（VSV）和甲型流感病毒（IAV）。使用基因沉默，ATP 1A 1蛋白被证明是至关重要的感染细胞与鼠肝炎病毒（MHV），猫传染性腹膜炎病毒（FIPV），VSV，但不是与IAV。缺乏ATP 1A 1不影响病毒与宿主细胞的结合，但导致MHV和VSV的进入受到抑制。一致的是，纳摩尔浓度的强心类固醇哇巴因和蟾蜍灵，这是已知的不影响Na+，K+-ATP酶的转运功能，抑制感染细胞与MHV，FIPV，中东呼吸综合征（MERS）-CoV，和VSV，但不IAV，当化合物存在于病毒接种。强心类固醇显示在早期阶段抑制MHV的进入，导致病毒体靠近细胞表面的积累，并因此减少融合。与感染早期阻断一致，CTS对VSV的抑制可被低pH休克绕过。当病毒进入后加入这些化合物时，病毒RNA复制不受影响。哇巴因的抗病毒作用可以通过添加不同的Src激酶抑制剂来减轻，这表明通过ATP 1A 1介导的Src信号传导在抑制CoV和VSV感染中起着至关重要的作用。重要的是，冠状病毒（CoVs）是动物和人类的重要病原体，最近出现的新的人畜共患病来源的CoVs证明了这一点。缺乏针对CoV感染的抗病毒药物。在本研究中，我们表明，ATP 1A 1亚基的Na+，K+-ATP酶，离子转运蛋白和信号转导，支持冠状病毒感染。通过基因沉默或低浓度的ATP 1A 1结合强心类固醇哇巴因和蟾蜍灵靶向ATP 1A 1导致在早期进入阶段抑制鼠，猫和MERS CoV的感染。对照病毒VSV的感染也受到抑制。由ATP 1A 1介导的Src信号在哇巴因和蟾蜍灵抑制病毒进入中起关键作用。这些结果表明，靶向Na+，K+-ATP酶使用强心类固醇，其中几个是FDA批准的化合物，可能是一个有吸引力的治疗方法对冠状病毒和VSV感染。
In addition to transporting ions, the multisubunit Na+,K+-ATPase also functions by relaying cardiotonic steroid (CTS)-binding-induced signals into cells. In this study, we analyzed the role of Na+,K+-ATPase and, in particular, of its ATP1A1 alpha subunit during coronavirus (CoV) infection. As controls, the vesicular stomatitis virus (VSV) and influenza A virus (IAV) were included. Using gene silencing, the ATP1A1 protein was shown to be critical for infection of cells with murine hepatitis virus (MHV), feline infectious peritonitis virus (FIPV), and VSV but not with IAV. Lack of ATP1A1 did not affect virus binding to host cells but resulted in inhibited entry of MHV and VSV. Consistently, nanomolar concentrations of the cardiotonic steroids ouabain and bufalin, which are known not to affect the transport function of Na+,K+-ATPase, inhibited infection of cells with MHV, FIPV, Middle East respiratory syndrome (MERS)-CoV, and VSV, but not IAV, when the compounds were present during virus inoculation. Cardiotonic steroids were shown to inhibit entry of MHV at an early stage, resulting in accumulation of virions close to the cell surface and, as a consequence, in reduced fusion. In agreement with an early block in infection, the inhibition of VSV by CTSs could be bypassed by low-pH shock. Viral RNA replication was not affected when these compounds were added after virus entry. The antiviral effect of ouabain could be relieved by the addition of different Src kinase inhibitors, indicating that Src signaling mediated via ATP1A1 plays a crucial role in the inhibition of CoV and VSV infections.IMPORTANCECoronaviruses (CoVs) are important pathogens of animals and humans, as demonstrated by the recent emergence of new human CoVs of zoonotic origin. Antiviral drugs targeting CoV infections are lacking. In the present study, we show that the ATP1A1 subunit of Na+,K+-ATPase, an ion transporter and signaling transducer, supports CoV infection. Targeting ATP1A1 either by gene silencing or by low concentrations of the ATP1A1-binding cardiotonic steroids ouabain and bufalin resulted in inhibition of infection with murine, feline, and MERS-CoVs at an early entry stage. Infection with the control virus VSV was also inhibited. Src signaling mediated by ATP1A1 was shown to play a crucial role in the inhibition of virus entry by ouabain and bufalin. These results suggest that targeting the Na+,K+-ATPase using cardiotonic steroids, several of which are FDA-approved compounds, may be an attractive therapeutic approach against CoV and VSV infections.