The Role of Ebola Virus VP24 Nuclear Trafficking Signals in Infectious Particle Production.

Ebola virus (EBOV) viral protein 24 (VP24) interacts with importin-α (IMPA) nuclear transport proteins to inhibit interferon signaling. VP24 also contains a nuclear export signal (NES). The capacity to bind IMPA and the presence of a NES suggest that VP24 traffics to the nucleus. However, this had not been demonstrated in the context of EBOV replication. In addition, whether these signals influence virus replication beyond effects on interferon responses has been unclear. Here, we demonstrate that VP24 traffics to the nucleus via IMPA during EBOV infection and in the context of an EBOV transcription and replication competent virus-like particle (trVLP) assay. Using the trVLP system, we also confirmed that VP24 possesses a functional nuclear export signal (NES). Mutations disrupting VP24-IMPA interaction or nuclear export reduced or abrogated trVLP infectivity, respectively. VP24 is known to be necessary for viral nucleocapsid maturation. The VP24 IMPA interface mutants yielded shortened nucleocapsids while VP24 NES mutations led to loss of nucleocapsid formation. Interestingly, VP24 mutants with intact NES function but altered sequences near the NES remain incapable of producing nucleocapsids. Together, these data demonstrate VP24 undergoes nuclear trafficking and reveals additional roles for both the IMPA interaction interface and the NES in nucleocapsid assembly.