Macroautophagy Proteins Assist Epstein Barr Virus Production and Get Incorporated Into the Virus Particles

• Macroautophagic membranes are stabilized during lytic EBV replication.
• Inhibition of macroautophagic membrane formation reduces EBV production.
• Stimulation of macroautophagic membrane formation boosts EBV production.
• Without macroautophagic membranes EBV DNA accumulates in the cytosol.
• Macroautophagic membranes get incorporated into EBV particles.

Epstein Barr virus (EBV) persists as a latent herpes virus infection in the majority of the adult human population. The virus can reactivate from this latent infection into lytic replication for virus particle production. Here, we report that autophagic membranes, which engulf cytoplasmic constituents during macroautophagy and transport them to lysosomal degradation, are stabilized by lytic EBV replication in infected epithelial and B cells. Inhibition of autophagic membrane formation compromises infectious particle production and leads to the accumulation of viral DNA in the cytosol. Vice versa, pharmacological stimulation of autophagic membrane formation enhances infectious virus production. Atg8/LC3, an essential macroautophagy protein and substrate anchor on autophagic membranes, was found in virus preparations, suggesting that EBV recruits Atg8/LC3 coupled membranes to its envelope in the cytosol. Our data indicate that EBV subverts macroautophagy and uses autophagic membranes for efficient envelope acquisition during lytic infection.