DDX3X Promotes Rotavirus Infection and Serves as an Antiviral Target

Rotavirus (RV) is a significant zoonotic pathogen primarily causing severe diarrheal disease in humans and animals, posing substantial risks to global public health and livestock industries. VP4 is one of the outer capsid proteins of RV and plays a crucial role in RV attachment and internalization. Additionally, it is also involved in replication and immune responses during RV infection; however, related studies are still limited. Here, a comprehensive analysis of the RV VP4 interactome was conducted, and DDX3X, one of six DExD/H helicase family members identified as interacting with VP4, potentially plays a crucial role in RV infection. Silencing DDX3X inhibits RV infection, whereas its overexpression facilitates RV infection. Further research demonstrated that VP4 interacts with DDX3X and the enzymatic activity of DDX3X was found to contribute to promote RV replication. Additionally, a drug screening study based on the VP4 interactome identified RK-33, a potent inhibitor of DDX3X, as the most effective candidate compound for inhibiting RV. In conclusion, VP4 interacts with DDX3X and the enzymatic activity of DDX3X is crucial for RV replication. The DDX3X inhibitor RK-33 exhibits significant inhibitory effects on RV infection. This study highlights the important roles of DDX3X in RV infection, offering potential candidate drugs for RV and expanding our understanding of its mechanisms.