title: Focal adhesion kinase is involved in rabies virus infection through its interaction with viral phosphoprotein P.
abstract: The rabies virus (RABV) phosphoprotein P is a multifunctional protein: it plays an essential role in viral transcription and replication, and in addition, RABV P has been identified as an interferon antagonist. Here, a yeast two-hybrid screen revealed that RABV P interacts with the focal adhesion kinase (FAK). The binding involved the 106-to-131 domain, corresponding to the dimerization domain of P and the C-terminal domain of FAK containing the proline-rich domains PRR2 and PRR3. The P-FAK interaction was confirmed in infected cells by coimmunoprecipitation and colocalization of FAK with P in Negri bodies. By alanine scanning, we identified a single mutation in the P protein that abolishes this interaction. The mutant virus containing a substitution of Ala for Arg in position 109 in P (P.R109A), which did not interact with FAK, is affected at a posttranscriptional step involving protein synthesis and viral RNA replication. Furthermore, FAK depletion inhibited viral protein expression in infected cells. This provides the first evidence of an interaction of RABV with FAK that positively regulates infection.
  
  
  OBJECTIVE
  Rabies virus exhibits a small genome that encodes a limited number of viral proteins. To maintain efficient virus replication, some of them are multifunctional, such as the phosphoprotein P. We and others have shown that P establishes complex networks of interactions with host cell components. These interactions have revealed much about the role of P and about host-pathogen interactions in infected cells. Here, we identified another cellular partner of P, the focal adhesion kinase (FAK). Our data shed light on the implication of FAK in RABV infection and provide evidence that P-FAK interaction has a proviral function.
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France.
authors.forename: Baptiste
authors.initials: B
authors.lastname: Fouquet
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France.
authors.forename: Jovan
authors.initials: J
authors.lastname: Nikolic
authors.affiliation: Institut Pasteur, Unit Lyssavirus Dynamics and Host Adaptation, Paris, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Larrous
authors.affiliation: Institut Pasteur, Unit Lyssavirus Dynamics and Host Adaptation, Paris, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Bourhy
authors.affiliation: Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Wirblich
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France danielle.blondel@vms.cnrs-gif.fr lagaudriere@vms.cnrs-gif.fr.
authors.forename: Cécile
authors.initials: C
authors.lastname: Lagaudrière-Gesbert
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France danielle.blondel@vms.cnrs-gif.fr lagaudriere@vms.cnrs-gif.fr.
authors.forename: Danielle
authors.initials: D
authors.lastname: Blondel
mesh_terms: D000818:Animals; D002460:Cell Line; D004252:DNA Mutational Analysis; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D007181:Inclusion Bodies, Viral / Q000737:chemistry / Q000821:virology; D018613:Microscopy, Confocal; D018832:Molecular Chaperones; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011820:Rabies virus / Q000502:physiology*; 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; D015678:Viral Structural Proteins; D051416:Focal Adhesion Protein-Tyrosine Kinases