Ruminant Diseases Research Center, College of Life Sciences, Shandong Normal University, 88 East of Wen-hua Road, Jinan, 250014, Shandong, PR China.
Virus Genes. 2021 Jun;57(3):250-257. doi: 10.1007/s11262-021-01833-0. Epub 2021 Feb 26.
Vesicular stomatitis virus (VSV) is an archetypal member of Mononegavirales which causes important diseases in cattle, horses and pigs. The matrix protein (M) of VSV plays critical roles in the replication, assembly/budding and pathogenesis of VSV. To further investigate the role of M during viral growth, we used a two-hybrid system to screen for host factors that interact with the M protein. Here, NADH: ubiquinone oxidoreductase complex assembly factor 4 (Ndufaf4) was identified as an M-binding partner, and this interaction was confirmed by yeast cotransformation and GST pulldown assays. The globular domain of M was mapped and shown to be critical for the M-Ndufaf4 interaction. Two double mutations (E156A/H157A, D180A/E181A) in M impaired the M-Ndufaf4 interaction. Overexpression of Ndufaf4 inhibited VSV propagation, and knockdown of Ndufaf4 by short hairpin RNA (shRNA) markedly promoted VSV replication. Finally, we also demonstrate that the anti-VSV effect of Ndufaf4 is independent of activation of the type I IFN response. These results indicated that Ndufaf4 might exploit other mechanisms to affect VSV replication. In summary, we identify Ndufaf4 as a potential target for the inhibition of VSV propagation. These results provided further insight into the study of VSV pathogenesis.
水疱性口炎病毒（VSV）是 Mononegavirales 的典型成员，会导致牛、马和猪的重要疾病。VSV 的基质蛋白（M）在病毒的复制、组装/出芽和发病机制中起着关键作用。为了进一步研究 M 在病毒生长过程中的作用，我们使用双杂交系统筛选与 M 蛋白相互作用的宿主因子。在这里，NADH：泛醌氧化还原酶复合物组装因子 4（Ndufaf4）被鉴定为 M 结合伴侣，酵母共转化和 GST 下拉测定证实了这种相互作用。M 的球状结构域被定位并显示对 M-Ndufaf4 相互作用至关重要。M 中的两个双突变（E156A/H157A、D180A/E181A）破坏了 M-Ndufaf4 相互作用。Ndufaf4 的过表达抑制了 VSV 的繁殖，短发夹 RNA（shRNA）的敲低则显著促进了 VSV 的复制。最后，我们还证明了 Ndufaf4 对 VSV 的抗病毒作用不依赖于 I 型干扰素反应的激活。这些结果表明，Ndufaf4 可能利用其他机制影响 VSV 的复制。总之，我们将 Ndufaf4 鉴定为抑制 VSV 繁殖的潜在靶标。这些结果为研究 VSV 的发病机制提供了进一步的见解。
