Classical swine fever virus (CSFV) nonstructural protein NS5A is a multifunctional protein functioning in regulation of viral genome replication, protein translation and assembly by interaction with viral or host proteins. Here, heat shock protein 27 (Hsp27) has been identified as a novel binding partner of NS5A by using His tag “pull down” coupled with shotgun LC-MS/MS, with interaction of both proteins further confirmed by co-immunoprecipitation and laser confocal assays. In PK-15 cells, silencing of Hsp27 expression by siRNA enhanced CSFV replication, and upregulation of Hsp27 inhibited viral proliferation. Additionally, we have shown that overexpression of Hsp27 increased NF-κB signaling induced by TNFα. Blocking NF-κB signaling in PK-15 cells overexpressing Hsp27 by ammonium pyrrolidinedithiocarbamate (PDTC) eliminated the inhibition of CSFV replication by Hsp27. These findings clearly demonstrate that the inhibition of CSFV replication by Hsp27 is mediated via the NF-κB signaling pathway.

古典猪瘟病毒（CSFV）非结构蛋白NS5A是一种多功能蛋白，可通过与病毒或宿主蛋白相互作用来调节病毒基因组复制，蛋白翻译和装配。在这里，热休克蛋白27（Hsp27）已通过使用His标签“下拉”结合shot弹枪LC-MS / MS被鉴定为NS5A的新型结合伴侣，并且通过共免疫沉淀和激光共聚焦进一步证实了这两种蛋白的相互作用分析。在PK-15细胞中，通过siRNA沉默Hsp27表达可增强CSFV复制，并且Hsp27的上调可抑制病毒增殖。此外，我们已经表明，Hsp27的过表达增加了TNFα诱导的NF-κB信号传导。吡咯烷二硫代氨基甲酸铵（PDTC）阻断过表达Hsp27的PK-15细胞中的NF-κB信号消除了Hsp27对CSFV复制的抑制作用。这些发现清楚地表明，Hsp27对CSFV复制的抑制作用是通过NF-κB信号传导途径介导的。