title: Interaction of rabies virus P-protein with STAT proteins is critical to lethal rabies disease.
abstract: BACKGROUND
  Rabies virus (RABV) causes rabies disease resulting in >55,000 human deaths/year. The multifunctional RABV P-protein has essential roles in genome replication, and forms interactions with cellular STAT proteins that are thought to underlie viral antagonism of interferon-dependent immunity. However, the molecular details of P-protein-STAT interaction, and its importance to disease are unresolved.
  
  
  METHODS
  Studies were performed using sequence/structure analysis, mutagenesis, immunoprecipitation, luciferase and qRT-PCR-based signaling assays, confocal microscopy and reverse genetics/in vivo infection.
  
  
  RESULTS
  We identified a hydrophobic pocket of the P-protein C-terminal domain as critical to STAT-binding/antagonism. This interface was found to be functionally and spatially independent of the region responsible for N-protein interaction, which is critical to genome replication. Based on these findings, we generated the first mutant RABV lacking STAT-association. Growth of the virus in vitro was unimpaired, but it lacked STAT-antagonist function and was highly sensitive to interferon. Importantly, growth of the virus was strongly attenuated in brains of infected mice, producing no major neurological symptoms, compared with the invariably lethal wild-type virus.
  
  
  CONCLUSIONS
  These data represent direct evidence that P-protein-STAT interaction is critical to rabies, and provide novel insights into the mechanism by which RABV coordinates distinct functions in interferon antagonism and replication.
authors.affiliation: Viral Pathogenesis Laboratory.
authors.forename: Linda
authors.initials: L
authors.lastname: Wiltzer
authors.forename: Kazuma
authors.initials: K
authors.lastname: Okada
authors.forename: Satoko
authors.initials: S
authors.lastname: Yamaoka
authors.forename: Florence
authors.initials: F
authors.lastname: Larrous
authors.forename: Henna Veera
authors.initials: HV
authors.lastname: Kuusisto
authors.forename: Makoto
authors.initials: M
authors.lastname: Sugiyama
authors.forename: Danielle
authors.initials: D
authors.lastname: Blondel
authors.forename: Hervé
authors.initials: H
authors.lastname: Bourhy
authors.forename: David Andrew
authors.initials: DA
authors.lastname: Jans
authors.forename: Naoto
authors.initials: N
authors.lastname: Ito
authors.forename: Gregory William
authors.initials: GW
authors.lastname: Moseley
mesh_terms: D000818:Animals; D002460:Cell Line; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D016679:Genome, Viral; D006801:Humans; D007372:Interferons / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D018832:Molecular Chaperones; D009154:Mutation; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011818:Rabies / Q000821:virology*; D011820:Rabies virus / Q000378:metabolism*; D050791:STAT Transcription Factors / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015678:Viral Structural Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D018832:Molecular Chaperones; C103961:P phosphoprotein, Rabies virus; D010750:Phosphoproteins; D050791:STAT Transcription Factors; D015678:Viral Structural Proteins; D007372:Interferons
keywords: Duvenhage virus; Viral disease; immune evasion; interferon; interferon antagonist; lyssavirus; pathogenicity; rabies virus; replication; signal transducers and activators of transcription