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.

水泡性口炎病毒 (VSV) 是单核病毒属的原型成员，可引起牛、马和猪的重要疾病。VSV 的基质蛋白 (M) 在 VSV 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，我们使用了一个双杂交系统来筛选与 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 结合伙伴，这种相互作用通过酵母共转化和 GST 下拉测定得到证实。M 的球状结构域被绘制并显示对 M-Ndufaf4 相互作用至关重要。M 中的两个双突变（E156A/H157A、D180A/E181A）削弱了 M-Ndufaf4 的相互作用。Ndufaf4 的过表达抑制了 VSV 的传播，和通过短发夹 RNA (shRNA) 敲低 Ndufaf4 显着促进了 VSV 复制。最后，我们还证明 Ndufaf4 的抗 VSV 作用与 I 型 IFN 反应的激活无关。这些结果表明 Ndufaf4 可能利用其他机制来影响 VSV 复制。总之，我们将 Ndufaf4 确定为抑制 VSV 传播的潜在目标。这些结果为 VSV 发病机制的研究提供了进一步的见解。