Neddylation of M1 negatively regulates the replication of influenza A virus
Neddylation of M1 negatively regulates the replication of influenza A virus
复制标题
M1 的 Neddylation 负向调节甲型流感病毒的复制
DOI：
10.1099/jgv.0.001503
复制
发表时间：
2020-01-01
影响因子：
3.8
通讯作者：
Ye, Xin
中科院分区：
文献类型：
--
作者：
Li, Yucen;Chai, Wenjia;Ye, Xin
翻译后修饰在病毒复制中起着关键作用。我们曾报道过A型流感病毒PB 2的奈迪化可以抑制病毒的复制。然而，我们发现NEDD 8过表达仍然可以抑制PB 2 K699 R突变病毒的复制，这意味着其他病毒蛋白可以被neddylated。在本研究中，我们发现IAV的M1也可以被NEDD 8修饰。我们发现E3连接酶HDM 2显著促进M1 neddylation。此外，我们确定M1 K187为主要的neddylation位点。我们产生了IAV M1 K187 R突变体（WSN-M1 K187 R），并比较了野生型和突变型病毒在Madin-Darby犬肾（MDCK）细胞中的生长。数据显示，WSN-M1 K187 R的复制比野生型WSN更有效。更重要的是，我们观察到NEDD 8的过表达比WSN-M1 K187 R更有效地抑制了野生型WSN的复制。此外，我们发现neddylation缺陷型M1突变体（M1 K187 R）具有比野生型M1更长的半衰期，表明M1的neddylation降低了稳定性。然后我们进行了病毒感染试验，发现WSN-M1 K187 R在小鼠中表现出比野生型WSN更大的毒力，表明M1的neddylation减少了IAV在体内的复制。总之，我们发现HDM 2对M1的neddylation负调节M1的稳定性，这反过来又抑制病毒复制。
Post-translational modification plays a critical role in viral replication. Previously we reported that neddylation of PB2 of influenza A virus (IAV) can inhibit viral replication. However, we found that NEDD8 overexpression can still inhibit the replication of PB2 K699R mutant viruses, implying that other viral protein(s) can be neddylated. In this study, we revealed that M1 of IAV can also be modified by NEDD8. We found that the E3 ligase HDM2 significantly promotes M1 neddylation. Furthermore, we identified M1 K187 as the major neddylation site. We generated an IAV M1 K187R mutant (WSN-M1 K187R) and compared the growth of wild-type and mutant viruses in Madin-Darby canine kidney (MDCK) cells. The data showed that the replication of WSN-M1 K187R was more efficient than that of wild-type WSN. More importantly, we observed that overexpression of NEDD8 inhibited the replication of the wild-type WSN more effectively than that of WSN-M1 K187R. In addition, we found that the neddylation-deficient M1 mutant (M1 K187R) had a longer half-life than that of wild-type M1, indicating that the neddylation of M1 reduces stability. Then we performed a viral infection assay and found that WSN-M1 K187R exhibited greater virulence in mice than wild-type WSN, suggesting that the neddylation of M1 reduced IAV replication in vivo. In conclusion, we uncovered that neddylation of M1 by HDM2 negatively regulates the stability of M1, which in turn inhibits viral replication.