Oligomerization of Ebola virus VP30 is essential for viral transcription and can be inhibited by a synthetic peptide.
Transcription of Ebola virus (EBOV)-specific mRNA is driven by the nucleocapsid proteins NP, VP35, and L. This process is further dependent on VP30, an essential EBOV-specific transcription factor. The present study addresses the self-assembly of VP30 and the functional significance of this process for viral transcription and propagation. Essential for oligomerization of VP30 is a region spanning amino acids 94-112. Within this region a cluster of four leucine residues is of critical importance. Mutation of only one of these leucine residues resulted in oligomerization-deficient VP30 molecules that were no longer able to support EBOV-specific transcription. The essential role of homo-oligomerization for the function of VP30 was further corroborated by the finding that mixed VP30 oligomers consisting of VP30 and transcriptionally inactive VP30 mutants were impaired in their ability to support EBOV transcription. The dominant negative effect of these VP30 mutants was dependent on their ability to bind to VP30. The oligomerization of VP30 could be dose dependently inhibited by a 25-mer peptide (E30pep-wt) derived from the presumed oligomerization domain (IC50,1 mum). A control peptide (E30pep-3LA), in which three leucines were changed to alanine, had no inhibitory effect. Thus, E30pep-wt seemed to bind efficiently to VP30 and consequently blocked the oligomerization of the protein. When E30pep-wt was transfected into EBOV-infected cells, the peptide inhibited viral replication suggesting that inhibition of VP30 oligomerization represents a target for EBOV antiviral drugs.
Hartlieb
Bettina
B
Institut für Virologie der Philipps-Universität Marburg, Robert-Koch-Strasse 17, 35037 Marburg, Germany.
Modrof
Jens
J
Mühlberger
Elke
E
Klenk
Hans-Dieter
HD
Becker
Stephan
S
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001665:Binding Sites; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D029043:Ebolavirus / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D007930:Leucine; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000494:pharmacology; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*
D010455:Peptides; D014157:Transcription Factors; C479897:VP30 protein, ebola virus; D014764:Viral Proteins; D007930:Leucine