ABSTRACT
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.
Subject(s)
Ebolavirus/metabolism , Protein Phosphatase 2/metabolism , Transcription Factors/genetics , Transcription, Genetic/genetics , Viral Proteins/genetics , Virus Replication/genetics , Animals , Cell Line, Tumor , Chlorocebus aethiops , Ebolavirus/genetics , HEK293 Cells , HeLa Cells , Humans , Nucleoproteins , Phosphorylation , Protein Interaction Domains and Motifs/genetics , Protein Phosphatase 2/antagonists & inhibitors , RNA, Viral/metabolism , Vero CellsABSTRACT
Nanocomposite systems comprised of a poly(ethylene vinyl acetate) (EVA) matrix and carbon black (CB) or graphene nanoplatelets (GNPs) were used to investigate conductivity and crystallisation dynamics using a commercially relevant melt-state mixing process. Crystallisation kinetics and morphology, as investigated by DSC and SEM, turn out to depend on the interplay of (i) the interphase interactions between matrix and filler, and (ii) the degree of filler agglomeration. For the GNP-based systems, an almost constant conductivity value was observed for all compositions upon cooling, something not observed for the CB-based compositions. These conductivity changes reflect structural and morphological changes that can be associated with positive and negative thermal expansion coefficients. GNP-based systems were observed to exhibit a percolation threshold of approximately 2.2 vol%, lower than the 4.4 vol% observed for the CB-based systems.