Interaction between the yellow fever virus nonstructural protein NS3 and the host protein Alix contributes to the release of infectious particles

The ESCRT (endosomal sorting complex required for transport) machinery normally executes cargo sorting and internalization during multivesicular body biogenesis, but is also utilized by several enveloped viruses to facilitate their budding from cellular membranes. Although the mechanisms of flavivirus infectious particle assembly and release are poorly understood, the nonstructural protein NS3 has been reported to have an essential role via an undescribed mechanism. Here, we shed light on the role of NS3 by connecting it to the host factor Alix, a protein intimately connected with the ESCRT machinery. We demonstrate that NS3 and Alix interact and show that dominant negative versions of Alix inhibit YFV release. Furthermore, we show that NS3 supplied in trans rescues this effect. We propose that the interaction between NS3 and Alix contributes to YFV release.