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Influenza infection modulates vesicular trafficking and induces Golgi complex disruption

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Abstract

Influenza A virus (IFV) replicates its genome in the nucleus of infected cells and uses the cellular protein transport system for genome trafficking from the nucleus to the plasma membrane. However, many details of the mechanism of this process, and its relationship to subsequent cytoplasmic virus trafficking, have not been elucidated. We examined the effect of nuclear transport inhibitors Leptomycin B (LB), 5,6 dichloro-1-β-d-ribofuranosyl-benzimidazole (DRB), the vesicular transport inhibitor Brefeldin A (BFA), the caspase inhibitor ZWEHD, and microtubule inhibitor Nocodazole (NOC) on virus replication and intracellular trafficking of viral nucleoprotein (NP) from the nucleus to the ER and Golgi. Also, we carried out complementary studies to determine the effect of IFV on intracellular membranes. Inhibition of the CRM1 and TAP-P15 nuclear transport pathways by DRB and LB blocked completely the export of virus. Inhibition of vesicular trafficking by BFA, NOC, and ZWEHD also affected influenza infection. Interestingly, IFV infection induced fragmentation of the Golgi complex resulting in diffuse distribution of large and small vesicles throughout the cytoplasm. Live-cell microscopy revealed expansion of Golgi localization signals indicating progressive dispersion of Golgi positive structures, resulting in the disassembly of the Golgi ribbon structure. Other vesicular components (Rab1b, ARF1 and GBF1) were also found to be required for IFV infection. Furthermore, the exact step at which IFV infection disrupts vesicle trafficking was identified as the ER-Golgi intermediate compartment. These findings suggest that IFV NP is trafficked from the nucleus via the CRM1 and TAP pathways. IFV modulates vesicular trafficking inducing disruption of the Golgi complex. These studies provide insight on the ways in which IFV affects intracellular trafficking of different host proteins and will facilitate identification of useful pharmaceutical targets to abrogate virus replication.

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Acknowledgments

We thank Adolfo Garcia Sastre, Monty Krieger, Nihal Altan-Bonnet, Mathias P. Machner for providing NS1-GFP virus, ldlf cells, ARF1, GBF1, Rab1b plasmids.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Tulane School of Medicine, Tulane University, New Orleans, LA, USA

    Kerstin Honer Zu Bentrup & Thomas G. Voss

  2. Division of Microbiology, Tulane National Primate Research Center, Covington, LA, USA

    Vibha Yadav & Antonito T. Panganiban

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  1. Vibha Yadav
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  3. Kerstin Honer Zu Bentrup
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Correspondence to Vibha Yadav.

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Yadav, V., Panganiban, A.T., Honer Zu Bentrup, K. et al. Influenza infection modulates vesicular trafficking and induces Golgi complex disruption. VirusDis. 27, 357–368 (2016). https://doi.org/10.1007/s13337-016-0347-3

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  • DOI: https://doi.org/10.1007/s13337-016-0347-3

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