The Lymphocytic Choriomeningitis Virus Matrix Protein PPXY Late Domain Drives the Production of Defective Interfering Particles

Arenaviruses cause severe diseases in humans but establish asymptomatic, lifelong infections in rodent reservoirs. Persistently-infected rodents harbor high levels of defective interfering (DI) particles, which are thought to be important for establishing persistence and mitigating virus-induced cytopathic effect. Little is known about what drives the production of DI particles. We show that neither the PPXY late domain encoded within the lymphocytic choriomeningitis virus (LCMV) matrix protein nor a functional endosomal sorting complex transport (ESCRT) pathway is absolutely required for the generation of standard infectious virus particles. In contrast, DI particle release critically requires the PPXY late domain and is ESCRT-dependent. Additionally, the terminal tyrosine in the PPXY motif is reversibly phosphorylated and our findings indicate that this posttranslational modification may regulate DI particle formation. Thus we have uncovered a new role for the PPXY late domain and a possible mechanism for its regulation.
Author Summary Arenaviruses cause severe and often fatal diseases in humans yet typically establish lifelong, asymptomatic infections in their rodent reservoirs. Several families of enveloped RNA viruses, including the arenaviruses, encode short amino acid motifs, called late domains, to hijack host proteins in the endosomal sorting complex required for transport (ESCRT) to drive the release of virus particles from the host cell’s outer membrane. Many late domain-containing viruses produce defective interfering (DI) particles in addition to standard, infectious virus. DI particles cannot self-replicate but interfere with the production of infectious virus and mitigate virus-induced cytopathic effect. Arenaviruses such as lymphocytic choriomeningitis virus (LCMV) generate high levels of DI particles, yet, the mechanism that drives their formation is not known. We show that LCMV’s only encoded late domain, PPXY, and a functional ESCRT pathway are critical for DI particle production, but in contrast, are not absolutely required for infectious virus production. We also demonstrate that the LCMV PPXY late domain is phosphorylated and that this modification may regulate DI particle production. In summary, we have discovered a new and unexpected role for a viral late domain in selectively driving the production of DI particles independently of standard infectious virus particles.