SNAP23 is essential for germination of EV-D68 replication organelles

Picornaviruses rearrange host cell membranes to facilitate their own replication. Here we investigate the Qbc SNARE, SNAP23, which is found at the plasma membrane and plays roles in exocytosis. We found that knockdown of SNAP23 expression inhibits virus replication but not release from cells. Knocking down SNAP23 inhibits viral RNA replication and synthesis of structural proteins. Normal cellular levels of SNAP23 are required for an early step in virus production, prior to or at the stage of virus RNA replication. We report that SNAP23 knockdown generates large, electron-light structures, and that infection of cells with these structures does not alter them, and those cells fail to generate viral RNA replication sites. We suggest that SNAP23 may play a role in maintaining membranes and lipids needed for generating virus replication organelles. Further investigation is needed to determine the precise role of this crucial SNARE protein in EV-D68 replication. •The cellular Qbc SNARE protein SNAP23 relocalizes from the plasma membrane to the cytosol after infection with EV-D68.•SNAP23 knockdown induces large, cytosolic, electron-light vacuoles. These vacuoles are still intact after infection.•Loss of SNAP23 affects viral RNA replication and generation of viral replication organelles.•Loss of SNAP23 leads to resources needed for building replication organelles being diverted into large vacuolar structures.