title: Molecular association of herpes simplex virus type 1 glycoprotein E with membrane protein Us9.
abstract: Herpes simplex virus type 1 (HSV-1) glycoprotein E (gE), glycoprotein I (gI), and Us9 promote efficient anterograde axonal transport of virus from the neuron cytoplasm to the axon terminus. HSV-1 and PRV gE and gI form a heterodimer that is required for anterograde transport, but an association that includes Us9 has not been demonstrated. NS-gE380 is an HSV-1 mutant that has five amino acids inserted after gE residue 380, rendering it defective in anterograde axonal transport. We demonstrated that gE, gI and Us9 form a trimolecular complex in Vero cells infected with NS-gE380 virus in which gE binds to both Us9 and gI. We detected the complex using immunoprecipitation with anti-gE or anti-gI monoclonal antibodies in the presence of ionic detergents. Under these conditions, Us9 did not associate with gE in cells infected with wild-type HSV-1; however, using a nonionic detergent, TritonX-100, an association between Us9 and gE was detected in immunoprecipitates of both wild-type and NS-gE380-infected cells. The results suggest that the interaction between Us9 and gE is weak and disrupted by ionic detergents in wild-type infected cells. We postulate that the tight interaction between Us9 and gE leads to the anterograde spread defect in the NS-gE380 virus.
authors: Infectious Disease Division, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, 522F Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, PA, 19104-6073, USA. sawasthi@mail.med.upenn.edu.
authors: Sita
authors: S
authors: Awasthi
authors: Infectious Disease Division, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, 522F Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, PA, 19104-6073, USA.
authors: Harvey M
authors: HM
authors: Friedman
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D018259:Herpesvirus 1, Human / Q000502:physiology*; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins; D008074:Lipoproteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008074:Lipoproteins; D010750:Phosphoproteins; C109622:US9 protein, Human herpesvirus 1; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C044768:glycoprotein E, herpes simplex virus type 1; C064959:glycoprotein I, herpes simplex virus type 1