Cullin 3-mediated ubiquitination restricts enterovirus D68 replication and is counteracted by viral protease 3C.

UNLABELLED

Enterovirus D68 (EV-D68) has emerged as a significant threat to public health because of its association with respiratory illnesses and neurological complications, including acute flaccid myelitis. However, the molecular mechanisms underlying EV-D68 replication and pathogenesis remain unclear. Here, we revealed a novel interaction between EV-D68 and the host Cullin-RING E3 ligase system, specifically Cullin 3, which was reported to restrict viral replication. We initially demonstrated that proteasome inhibition enhanced EV-D68 replication, suggesting an important role for the ubiquitin-proteasome system in viral restriction. Cullin 3 was further identified as a key factor that inhibits EV-D68 replication, and the downregulation of its expression increased viral titers. Mechanistically, Cullin 3 was observed to target the viral capsid protein VP1 for ubiquitination and degradation. However, EV-D68 was determined to utilize its protease 3C to cleave Cullin 3 at the Q681 residue, thereby inhibiting E3 ligase activity and facilitating resistance to Cullin 3-mediated VP1 degradation. This study uncovered a host-virus arms race, wherein the ubiquitin-proteasome system of the host actively targets viral proteins for degradation, and viral proteases counteract this defense mechanism. Accordingly, these findings could lead to more effective antiviral treatments.

IMPORTANCE

The ubiquitin-proteasome system (UPS) is a critical cellular pathway involved in the regulation of protein stability and has been implicated in the regulation of viral infections. However, its role in EV-D68 infection has not been extensively explored. Our study proves that the host UPS, through the scaffold protein Cullin 3, can restrict EV-D68 replication, representing a previously unrecognized antiviral mechanism. Furthermore, we describe a viral strategy used to evade this host defense mechanism comprising Cullin 3 cleavage, which has broad implications for understanding virus-host interactions and could inform the development of novel therapeutic strategies against EV-D68 and other enteroviruses.