The Ebola Virus Nucleoprotein Recruits the Host PP2A-B56 Phosphatase to Activate Transcriptional Support Activity of VP30

Transcription of the Ebola virus genome depends on the viral transcription factor VP30 in its unphosphorylated form, but the underlying molecular mechanism of VP30 dephosphorylation is unknown. Here we show that the Ebola virus nucleoprotein (NP) recruits the host PP2A-B56 protein phosphatase through a B56-binding LxxIxE motif and that this motif is essential for VP30 dephosphorylation and viral transcription. The LxxIxE motif and the binding site of VP30 in NP are in close proximity, and both binding sites are required for the dephosphorylation of VP30. We generate a specific inhibitor of PP2A-B56 and show that it suppresses Ebola virus transcription and infection. This work dissects the molecular mechanism of VP30 dephosphorylation by PP2A-B56, and it pinpoints this phosphatase as a potential target for therapeutic intervention. [Display omitted] •The Ebola virus NP recruits the PP2A-B56 phosphatase through an LxxIxE binding motif•The LxxIxE motif in NP is required for VP30 dephosphorylation and viral transcription•A specific PP2A-B56 inhibitor impairs recombinant Ebola virus transcription and infection Transcription of the Ebola virus genome depends on the viral transcription factor VP30 in its unphosphorylated form. Kruse et al. show that the Ebola nucleoprotein recruits the host PP2A-B56 phosphatase through a B56-binding LxxIxE motif, leading to VP30 dephosphorylation and viral transcription.