Protein NS4A of ZIKV Inhibits Glycolytic Flux by Targeting Enolase-1

NS4A plays a role in forming the flavivirus replication complex, which inhibits apoptosis in host cells by inducing autophagy, thereby promoting viral replication. The host protein ENO1 interacts with NS4A, but the precise mechanism underlying this interaction and its role in viral replication remain unclear. In this study, we identified ZIKV NS4A1–73 as a key regulator of replication and infection cycles in both temporal and spatial dimensions. Through surface plasmon resonance (SPR) analysis, we demonstrated that ENO1 directly interacts with NS4A1–73. This critical binding inhibits the enzymatic activity of ENO1 and reduces cellular lactate and ATP production. Our findings suggest that ZIKV NS4A may effectively impede cellular metabolism by targeting the host factor ENO1, thus disrupting the glycolytic process. This insight could open new avenues for targeting ZIKV and similar viruses in therapeutic strategies.