Inhibitor of Sarco/Endoplasmic Reticulum Calcium-ATPase Impairs Multiple Steps of Paramyxovirus Replication
Inhibitor of Sarco/Endoplasmic Reticulum Calcium-ATPase Impairs Multiple Steps of Paramyxovirus Replication
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DOI：
10.3389/fmicb.2019.00209
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发表时间：
2019-02-13
影响因子：
5.2
通讯作者：
Barua, Sanjay
中科院分区：
文献类型：
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作者：
Kumar, Naveen;Khandelwal, Nitin;Barua, Sanjay
肌浆网/内质网钙ATP酶（SERCA）是一种膜结合的胞质酶，已知其调节肌浆网/内质网的钙摄取。在此，我们首次证明SERCA也可以调节病毒复制。用对细胞无毒浓度的SERCA特异性抑制剂（毒胡萝卜素）处理Vero细胞显著降低了小反刍兽疫病毒（PPRV）和纽卡斯尔病病毒（NDV）的复制。相反，SERCA的过表达挽救了毒胡萝卜素对病毒复制的抑制作用。PPRV和NDV感染可诱导Vero细胞SERCA基因表达，该表达可被Thapsigargin阻断。除了诱导增强的细胞质病灶形成外，毒胡萝卜素还显示出阻断病毒进入靶细胞以及病毒蛋白质的合成。此外，显示NDV在Vero细胞中长期传代（P）后获得对毒胡萝卜素的显著抗性。与P0和P70-对照相比，P70-Thapsigargin病毒的融合（F）蛋白在氨基酸残基104（E104 K）处显示出独特的突变，而在HN基因中未观察到Thapsigargin相关的突变。据我们所知，这是第一份描述SERCA病毒支持作用的报告，也是一份罕见的报告，表明即使在靶向细胞因子的抑制剂存在下，病毒也可能获得耐药性。
Sarco/endoplasmic reticulum calcium-ATPase (SERCA) is a membrane-bound cytosolic enzyme which is known to regulate the uptake of calcium into the sarco/endoplasmic reticulum. Herein, we demonstrate for the first time that SERCA can also regulate virus replication. Treatment of Vero cells with SERCA-specific inhibitor (Thapsigargin) at a concentration that is nontoxic to the cells significantly reduced Peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV) replication. Conversely, overexpression of SERCA rescued the inhibitory effect of Thapsigargin on virus replication. PPRV and NDV infection induced SERCA expression in Vero cells, which could be blocked by Thapsigargin. Besides inducing enhanced formation of cytoplasmic foci, Thapsigargin was shown to block viral entry into the target cells as well as synthesis of viral proteins. Furthermore, NDV was shown to acquire significant resistance to Thapsigargin upon long-term passage (P) in Vero cells. As compared to the P0 and P70-Control, the fusion (F) protein of P70-Thapsigargin virus exhibited a unique mutation at amino acid residue 104 (E104K), whereas no Thapsigargin-associated mutations were observed in HN gene. To the best of our knowledge, this is the first report describing the virus-supportive role of SERCA and a rare report suggesting that viruses may acquire resistance even in the presence of an inhibitor that targets a cellular factor.