The interaction between host Annexin A2 and viral Nsp9 is beneficial for replication of porcine reproductive and respiratory syndrome virus
The interaction between host Annexin A2 and viral Nsp9 is beneficial for replication of porcine reproductive and respiratory syndrome virus
复制标题
宿主Annexin A2与病毒Nsp9之间的相互作用有利于猪繁殖与呼吸综合征病毒的复制
DOI：
10.1016/j.virusres.2014.05.015
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发表时间：
2014-08-30
期刊：
影响因子：
5
通讯作者：
Weng, Changjiang
中科院分区：
文献类型：
--
作者：
Li, Jiangnan;Guo, Dongwei;Weng, Changjiang
非结构蛋白9(Nsp9)是猪繁殖与呼吸综合征病毒(PRRSV)的一种依赖RNA的RNA聚合酶(RdRp)，是PRRSV复制所必需的。然而，NSP9的结合伙伴尚未确定。在本研究中，利用酵母双杂交技术(Y2H)鉴定了7种寄主蛋白为Nsp9结合蛋白。其中，我们用Y2H、免疫共沉淀(Co-IP)、GST下拉和免疫荧光分析(IFA)证实了Nsp9与Annexin A2(ANXA2)的相互作用。我们发现只有全长ANXA2在体外才能与Nsp9结合，而在PRRSV感染的Marc-145细胞中，Nsp9与内源性ANXA2相互作用。此外，我们还发现Nsp9-ANXA2的相互作用被RNaseA处理部分降低。此外，PRRSV在ANXA2基因敲除的MARC-145细胞中的生长受到显著抑制。综上所述，这些结果表明Nsp9结合伙伴ANXA2有利于PRRSV的复制。(C)2014爱思唯尔B.V.保留所有权利。
Non-structural protein 9 (Nsp9), a RNA-dependent RNA polymerase (RdRp) of the porcine reproductive and respiratory syndrome virus (PRRSV), is necessary for PRRSV replication. However, the binding partners of Nsp9 have not been identified. In this study, seven host proteins were identified as Nsp9-binding proteins using Yeast two-hybrid (Y2H). Among of them, we confirmed the interaction of Nsp9 with Annexin A2 (ANXA2) using Y2H, Co-immunoprecipitation (Co-IP), GST pulldown and immunofluorescence assay (IFA). We found that only full-length ANXA2 could bind with Nsp9 in vitro and Nsp9 interacted with endogenous ANXA2 in PRRSV-infected MARC-145 cells. In addition, we found that the Nsp9-ANXA2 interaction was partially reduced by RNase A treatment. Furthermore, PRRSV growth was significantly hindered in ANXA2-knockdown MARC-145 cells. Taken together, these results indicate that Nsp9 binding partner ANXA2 is beneficial for PRRSV replication. (C) 2014 Elsevier B.V. All rights reserved.