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Molecular association of herpes simplex virus type 1 glycoprotein E with membrane protein Us9

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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.

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Acknowledgments

We thank Toni Minson, Gary Cohen and Roselyn Eisenberg for providing reagents. We also thank the University of Pennsylvania Confocal Microscopy Core facility for their assistance.

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  1. Infectious Disease Division, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, 522F Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, PA, 19104-6073, USA

    Sita Awasthi & Harvey M. Friedman

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  1. Sita Awasthi
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  2. Harvey M. Friedman
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Correspondence to Sita Awasthi.

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This study was funded by R01-AI104854 to HF.

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Authors SA and HF declare that they have no conflict of interest.

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Awasthi, S., Friedman, H.M. Molecular association of herpes simplex virus type 1 glycoprotein E with membrane protein Us9. Arch Virol 161, 3203–3213 (2016). https://doi.org/10.1007/s00705-016-3028-z

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