Prazoles Targeting Tsg101 Inhibit Release of Epstein-Barr Virus following Reactivation from Latency

Epstein-Barr virus (EBV) is a ubiquitous herpesvirus responsible for several diseases including cancers of lymphoid and epithelial cells. EBV-cancers typically exhibit viral latency; however, the production and release of EBV through its lytic phase is essential for cancer development. Antiviral agents that specifically target EBV production do not currently exist. Previously, we reported that the proton pump inhibitor Tenatoprazole, which blocks the interaction of ubiquitin with the ESCRT-1 factor Tsg101, inhibits production of several enveloped viruses, including EBV. Here, we show that three structurally distinct prazoles impair mature particle formation post-reactivation and identify the impact on stages of replication. The prazoles did not impair expression of lytic genes representative of the different kinetic classes but interfered with capsid maturation in the nucleus as well as virion transport from the nucleus. Replacement of endogenous Tsg101 with a mutant Tsg101 refractory to prazole-mediated inhibition rescued EBV release. These findings directly implicate Tsg101 in EBV nuclear egress, and identify prazoles as potential therapeutic candidates for conditions that rely on EBV replication such as chronic active EBV infection and post-transplant lymphoproliferative disorders. IMPORTANCE Production of virions is necessary for the ubiquitous Epstein-Barr virus (EBV) to persist in humans but in so doing, can set the stage for development of EBV-cancers in at-risk individuals. In our attempts to identify inhibitors of the EBV lytic phase, we previously found that a prazole proton pump inhibitor, known to block the interaction of ubiquitin with the ESCRT-1 factor Tsg101, blocks production of EBV. We now find that three structurally distinct prazoles impair maturation of EBV capsids and virion transport from the nucleus, and by interfering with Tsg101, prevent EBV release from lytically-active cells. Our findings not only implicate Tsg101 in EBV production but also identify widely-used prazoles as candidates to prevent development of post-transplant EBV-lymphomas.