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. IMPORTANCERabies 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.

Dysregulation of focal adhesion kinase upon Toxoplasma gondii infection facilitates parasite translocation across polarised primary brain endothelial cell monolayers

“…In contrast, Salmonella activates FAK to suppress autophagy and thereby promote bacterial survival in macrophages (Owen, Meyer, Bouton, & Casanova, 2014), a feature also recently described for T. gondii and discussed below (Portillo et al, 2017). Also, Hepatitis B virus activates FAK (Bouchard, Wang, & Schneider, 2006), and rabies virus interacts with FAK (Fouquet et al, 2015) to promote intracellular survival and replication. Thus, a number of pathogens are known to manipulate FAK functions.…”

Identification and characterization of the role of c-terminal Src kinase in dengue virus replication

“…Ebola virus has been shown to depend on the activity of tyrosine kinases at both entry and later stages of budding and release of virions 26 27 . The essential role of TK signaling in viral replication has been identified by a number of studies using multiple approaches such as pharmacological inhibitors of specific TKs, by expression of viruses harboring mutations in TK interaction regions or using siRNA screens against host TKs 12 28 29 30 31 32 . In most cases, viruses have been shown to activate the cytosolic TK signaling pathways to modulate the cellular environment in favor of virus replication.…”

Rabies Virus Infection Induces the Formation of Stress Granules Closely Connected to the Viral Factories

“…Recombinant viruses were recovered as described previously [ 62 , 63 ]. Briefly, N2A cells (10 6 cells) were transfected using lipofectamine 2000 (Invitrogen) with 0.85 μg of full-length prCVSN2C-P-mCherry, in addition to 0.4 μg pTIT-N, 0.2 μg pTIT-P, 0.2 μg pTIT-L and 0.15 μg pTIT-G, which encode respectively the N, P, L and G proteins of SAD-L16 rabies virus strain.…”