Autophagy Induced by the N-Terminus of the Classic Swine Fever Virus Nonstructural Protein 5A Protein Promotes Viral Replication.
Autophagy Induced by the N-Terminus of the Classic Swine Fever Virus Nonstructural Protein 5A Protein Promotes Viral Replication.
复制标题
猪瘟病毒非结构蛋白5A蛋白N端诱导自噬促进病毒复制
DOI：
10.3389/fmicb.2021.733385
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发表时间：
2021
影响因子：
5.2
通讯作者：
Wu Y
中科院分区：
文献类型：
--
作者：
Zhang C;Wang X;Sun J;Guo M;Zhang X;Wu Y
猪瘟病毒（CSFV）感染可诱导自噬，但其具体机制尚不清楚。猪瘟病毒非结构蛋白5A（NS 5A）是一种多磷酸化蛋白，具有调节病毒复制和宿主细胞免疫应答的多种功能。在此，我们证明了CSFV NS 5A可以诱导细胞自噬并促进病毒复制。在目前的研究中，我们发现NS 5A表达显著增加了自噬相关基因（ATG）的水平，包括轻链3（LC 3），ATG 5和Beclin 1;相反，通过Western印迹观察到P62/隔离体1（SQSTM 1）的降解。透射电子显微镜（TEM）观察到表达NS 5A的细胞中自噬样囊泡的数量也明显增加。此外，我们通过共聚焦免疫荧光分析观察到NS 5A和LC 3蛋白的共定位。NS 5A与自噬相关的LC 3蛋白的直接结合通过体内共免疫沉淀和体外GST下拉测定来证实。通过对NS 5A蛋白的分段和点突变研究，我们发现NS 5A蛋白的N端区域和81和92位氨基酸的磷酸化是诱导自噬所必需的。最后，我们证明了LC 3蛋白对CSFV复制具有积极作用。这些发现强调了先前未确定的自噬过程中NS 5A和LC 3之间的相互作用关系。此外，我们的研究揭示了CSFV NS 5A的新作用，特别是其N-末端氨基酸丝氨酸81和丝氨酸92，作为CSFV诱导的自噬的关键调节剂，并对扩展我们对CSFV-自噬相互作用的理解具有重要意义。
Although classic swine fever virus (CSFV) infection has been reported to induce autophagy, the specific induced mechanism remains unrevealed. Nonstructural protein 5A (NS5A) of CSFV is a multiphosphorylated protein with multiple functions to regulate viral replication and the host cell immune responses. Herein, we demonstrated that CSFV NS5A could induce cellular autophagy and promote viral replication. In the current study, we showed that NS5A expression significantly increased the levels of autophagy-related genes (ATGs), including light chain 3 (LC3), ATG5, and Beclin 1; conversely, degradation of P62/sequestosome 1 (SQSTM1) was observed by Western blotting. The number of autophagy-like vesicles was also obviously increased in NS5A-expressing cells, as analyzed by transmission electron microscopy (TEM). Furthermore, we observed the co-localization of the NS5A and LC3 proteins by confocal immunofluorescence analysis. Direct binding of NS5A to the autophagy-related LC3 protein was confirmed by coimmunoprecipitation in vivo and by a GST pulldown assay in vitro. Through segmentation and point mutation research on the NS5A protein, we found that the N-terminal region and the phosphorylation of amino acids 81 and 92 of the NS5A protein were essential for inducing autophagy. Finally, we demonstrated that the LC3 protein had a positive effect on CSFV replication. These findings emphasize a previously unascertained interaction relationship between NS5A and LC3 in the autophagy process. Furthermore, our research revealed a new role of CSFV NS5A, particularly its N-terminal amino acids serine 81 and serine 92, as a critical regulator of CSFV-induced autophagy and have significance for extending our understanding of the CSFV-autophagy interplay.
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影响因子：
6
作者：
Jassey, Alagie;Liu, Ching-Hsuan;Lin, Liang-Tzung
通讯作者：
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影响因子：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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