Multiple site SUMOylation of FMDV 3C protease and its negative role in viral replication

Authors: WU, XIANGJU - Shandong Academy Of Agricultural Sciences; HU, YUE - Shandong Academy Of Agricultural Sciences; SUI, CHAO - Shandong Academy Of Agricultural Sciences; PAN, LI - Chinese Academy Of Agricultural Sciences; YOO, DONGWAN - University Of Illinois; Miller, Laura; LEE, CHANGHEE - Gyeongsang National University; CONG, XIAOYAN - Shangdong Agricultural University; LI, JUNTONG - Shangdong Agricultural University; DU, YIJUN - Shandong Academy Of Agricultural Sciences; QI, JING - Shandong Academy Of Agricultural Sciences

Submitted to: Journal of Virology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/25/2022
Publication Date: 9/14/2022
Citation: Wu, X., Hu, Y., Sui, C., Pan, L., Li, F., Miller, L.C., Lee, C., Cong, X., Li, J., Du, Y., Qi, J. 2022. Multiple site SUMOylation of FMDV 3C protease and its negative role in viral replication. Journal of Virology. 96(17). Article e061222. https://doi.org/10.1128/jvi.00612-22.
DOI: https://doi.org/10.1128/jvi.00612-22

Interpretive Summary: Foot-and-mouth disease (FMD) is one of the most significant constraints to international trade in animals and animal products described by the World Organization for Animal Health. To develop control strategies, including antiviral approaches and vaccines, it is essential to gain a better understanding of the virus-host cell interactions. SUMOylation, the covalent linkage of a small ubiquitin-like protein to a variety of substrate proteins, has emerged as an extensively studied posttranslational modification (PTM) that plays important roles in diverse biological processes. In this study, four lysine residues were found to jointly determine the SUMOylation of FMDV 3C protease. In addition, we demonstrated that SUMOylation attenuates FMDV 3C function, including the cleavage ability, inhibitory effect of interferon signaling pathway and protein stability, which in turn results in a decrease in FMDV replication. Our findings indicate that SUMOylation of FMDV 3C serves as a host cell defense against virus replication. Further understanding of the cellular and molecular mechanisms driving this critical interaction should offer novel insights to design an effective strategy to control the dissemination of FMDV in animals.

Technical Abstract: Protein SUMOylation represents an important cellular process that regulates activities of numerous host proteins as well as of many invasive viral proteins. Foot-and-mouth disease virus (FMDV) is the first animal virus discovered. However, whether SUMOylation takes place during FMDV infection and what role it plays in FMDV pathogeny have not been investigated. In the present study, we demonstrated that SUMOylation suppresses FMDV replication by small interfering RNA (siRNA) transfection coupled with pharmaceutical inhibition of SUMOylation, which is confirmed by the high titer of FMDV 3C SUMOylation-deficient virus. Moreover, we provided evidence that four lysine residues Lys-51, 54, 110, 159 work together to conduct the SUMOylation of FMDV 3C protease. This is the first report of protein SUMOylation at multiple positions. Finally, we showed that SUMOylation attenuates the cleavage ability, the inhibitory effect of interferon signaling pathway and the protein stability of FMDV 3C, which correlates with a decrease in FMDV replication. Taken together, our results describe, for the first time, a cellular regulatory event that significantly restricts FMDV replication through the SUMOylation of 3C protease.