Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
Viruses. 2022 Feb 9;14(2):359. doi: 10.3390/v14020359.
Encephalomyocarditis virus can cause myocarditis and encephalitis in pigs and other mammals, thus posing a potential threat to public health safety. The 2A protein is an important virulence factor of EMCV. Previous studies have shown that the 2A protein may be related to the inhibition of apoptosis by virus, but its specific molecular mechanism is not clear. In this study, the 2A protein was expressed in in order to find interacting cell proteins. A pull down assay, coupled with mass spectrometry, revealed that the 2A protein possibly interacted with annexin A2. Co-immunoprecipitation assays and confocal imaging analysis further demonstrated that the 2A protein interacted with annexin A2 in cells. In reducing the expression of annexin A2 by siRNA, the ability of the 2A protein to inhibit apoptosis was weakened and the proliferation of EMCV was slowed down. These results suggest that annexin A2 is closely related to the inhibition of apoptosis by 2A. Furthermore, both RT-PCR and western blot results showed that the 2A protein requires annexin A2 interaction to inhibit apoptosis via JNK/c-Jun pathway. Taken together, our data indicate that the 2A protein inhibits apoptosis by interacting with annexin A2 via the JNK/c-Jun pathway. These findings provide insight into the molecular pathogenesis underlying EMCV infection.
脑心肌炎病毒可引起猪和其他哺乳动物的心肌炎和脑炎，因此对公共卫生安全构成潜在威胁。2A 蛋白是 EMCV 的重要毒力因子。先前的研究表明，2A 蛋白可能与病毒抑制细胞凋亡有关，但具体的分子机制尚不清楚。在本研究中，为了寻找相互作用的细胞蛋白，在 中表达了 2A 蛋白。下拉实验，结合质谱分析，显示 2A 蛋白可能与膜联蛋白 A2 相互作用。共免疫沉淀实验和共聚焦成像分析进一步表明，2A 蛋白在细胞内与膜联蛋白 A2 相互作用。通过 siRNA 降低膜联蛋白 A2 的表达，2A 蛋白抑制细胞凋亡的能力减弱，EMCV 的增殖速度减慢。这些结果表明，膜联蛋白 A2 与 2A 抑制细胞凋亡密切相关。此外，RT-PCR 和 Western blot 结果均表明，2A 蛋白通过 JNK/c-Jun 通路抑制细胞凋亡需要与膜联蛋白 A2 相互作用。综上所述，我们的数据表明，2A 蛋白通过 JNK/c-Jun 通路与膜联蛋白 A2 相互作用抑制细胞凋亡。这些发现为 EMCV 感染的分子发病机制提供了新的见解。
