ANKS4B Restricts Replication of Zika Virus by Downregulating the Autophagy
ANKS4B Restricts Replication of Zika Virus by Downregulating the Autophagy
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DOI：
10.3389/fmicb.2020.01745
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发表时间：
2020-07
影响因子：
5.2
通讯作者：
Quanshi Lin;Shili Zhou;Yanxia Huang;Zhiting Huo;Cancan Chen;Xin Luo;Junfang He;Chao Liu;
中科院分区：
文献类型：
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作者：
Quanshi Lin;Shili Zhou;Yanxia Huang;Zhiting Huo;Cancan Chen;Xin Luo;Junfang He;Chao Liu;
寨卡病毒（ZIKV）感染已成为全球健康的严重威胁，但目前尚无特效药物。在这项研究中，我们探讨了 ZIKV 与细胞蛋白、锚蛋白重复序列和含有 4b (ANKS4B) 的无菌基序结构域之间的关系。我们的数据显示，培养细胞和新生小鼠中 ANKS4B 的表达因 ZIKV 感染而下调。 ZIKV 感染后 ANKS4B 的减少是由两种肝细胞核因子 HNF1α 和 HNF4α 的减少引起的。通过CRISPR/Cas9基因编辑系统，我们分别在A549和Huh7细胞中生成了两个ANKS4B敲除（KO）细胞克隆。在ANKS4B-KO细胞中，包括病毒RNA、蛋白质和滴度在内的病毒复制水平显着增强，而ANKS4B的反式互补可逆转这种情况。 ANKS4B 不影响病毒进入步骤，但损害了 ZIKV 感染诱导的自噬。此外，我们的数据显示，抑制自噬导致 ANKS4B 充足和 ANKS4B 缺陷的细胞中 ZIKV 的复制水平相似，表明 ANKS4B 的抗病毒作用依赖于其对自噬的调节。因此，我们的工作将ANKS4B确定为ZIKV的新限制因子。
Infection of Zika virus (ZIKV) has become a severe threaten to global health while no specific drug is available. In this study, we explored the relationship between ZIKV and a cellular protein, ankyrin repeat and sterile motif domain containing 4b (ANKS4B). Our data revealed that the expression of ANKS4B in cultured cells and in neonatal mice was downregulated by ZIKV infection. The reduction of ANKS4B upon ZIKV infection was caused by decrease of two hepatocyte nuclear factors HNF1α and HNF4α. Through CRISPR/Cas9 gene editing system, we generated two ANKS4B knockout (KO) cell clones in A549 and Huh7 cells respectively. In the ANKS4B-KO cells, the viral replication levels including viral RNA, protein, and titer were significantly enhanced, which was reversed by trans-complementation of ANKS4B. ANKS4B did not affect the viral entry step, but impaired the autophagy induced by ZIKV infection. Furthermore, our data showed that inhibition of autophagy led to similar replication levels of ZIKV in ANKS4B-sufficient and ANKS4B-deficient cells, suggesting the antiviral effect of ANKS4B relied on its modulation on the autophagy. Therefore, our work identified ANKS4B as a new restriction factor of ZIKV.