Pan Wei, Song Deguang, He Wenqi, Lu Huijun, Lan Yungang, Li Hongli, Gao Feng, Zhao Kui
Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, Jilin University, 5333 Xi'an Road, Changchun, 130062, China.
Vet Microbiol. 2017 Dec;212:59-66. doi: 10.1016/j.vetmic.2017.10.021. Epub 2017 Nov 7.
The matrix protein of vesicular stomatitis virus (VSV) performs multiple functions during viral genome replication and virion production and is involved in modulating multiple host signaling pathways that favor virus replication. To perform numerous functions within infected cells, the M protein needs to recruit cellular partners. To better understand the role of M during VSV replication, we looked for interacting partners by using the two-hybrid system. The eukaryotic translation initiation factor 3, subunit i (eIF3i) was identified to be an M-binding partner, and this interaction was validated by GST pull-down and laser confocal assays. Through a mutagenesis analysis, we found that some mutants of M between amino acids 122 and 181 impaired but did not completely abolish the M-eIF3i interaction. Furthermore, the knockdown of eIF3i by RNA interference decreased viral replication and transcription in the early stages but led to increase in later stages. VSV transcription was increased at 4h post-infection but was not changed at 8 and 12h post-infection after the over-expression of eIF3i. Finally, we also demonstrated that VSV could inhibit the activity of Akt1 and that the knockdown of eIF3i inhibited the expression of the ISGs regulated by phospho-Akt1. These results indicated that eIF3i may affect VSV growth by regulating the host antiviral response in HeLa cells.
水泡性口炎病毒(VSV)的基质蛋白在病毒基因组复制和病毒粒子产生过程中发挥多种功能,并参与调节多种有利于病毒复制的宿主信号通路。为了在受感染细胞内执行众多功能,M蛋白需要招募细胞内的合作伙伴。为了更好地理解M蛋白在VSV复制过程中的作用,我们利用双杂交系统寻找相互作用的伙伴。真核翻译起始因子3亚基i(eIF3i)被鉴定为与M蛋白结合的伙伴,并且通过谷胱甘肽S-转移酶(GST)下拉实验和激光共聚焦实验验证了这种相互作用。通过诱变分析,我们发现M蛋白在氨基酸122和181之间的一些突变体削弱了但并未完全消除M-eIF3i相互作用。此外,通过RNA干扰敲低eIF3i在早期阶段会降低病毒复制和转录,但在后期阶段会导致增加。在感染后4小时,过表达eIF3i后VSV转录增加,但在感染后8小时和12小时没有变化。最后,我们还证明VSV可以抑制Akt1的活性,并且敲低eIF3i会抑制由磷酸化Akt-1调节的干扰素刺激基因(ISGs)的表达。这些结果表明,eIF3i可能通过调节HeLa细胞中的宿主抗病毒反应来影响VSV的生长。