Influenza infection modulates vesicular trafficking and induces Golgi complex disruption

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-b-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.

Immune responses in influenza A virus and human coronavirus infections: an ongoing battle between the virus and host

“…With the increasing accumulation of knowledge of molecular interactions between host cells and viruses, additional host molecules and normal biological processes [ 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66• , 67 , 68 , 69 ] were found to participate in the viral replication cycle (summarized in Table 2 ). To clarify the roles of these molecules and processes in virus infection, host genetic determinant screening [ 70 , 71 , 72 ], immunomics and Public Health Omics [ 73 ], host lipid omics [ 74 •• ] and characterization of the epigenetic landscape [ 75 ] were used to supplement conventional analyses.…”

“… [ 64 ] Vesicular trafficking IAV IAV infection modulates vesicular trafficking and induces Golgi complex disruption. [ 68 ] IAV enhances its propagation through modulating Annexin-A1 dependent endosomal trafficking. [ 51 ] IAV ribonucleoproteins modulate host recycling by competing with Rab11 effectors.…”

Golgi Structure and Function in Health, Stress, and Diseases

“…Several membrane structures including the Golgi are used by viruses as viral factories to replicate, concentrate, and assemble the viral genome and proteins into viral particles (Miller and Krijnse-Locker 2008;Netherton et al 2007;Salonen et al 2005). As a highly dynamic organelle, Golgi serves as a membrane scaffold for multiple viruses, including infectious hepatitis C virus, enteroviruses, poliovirus, foot-and-mouth-disease virus, dengue virus, coronavirus, Kunjin virus, tick-borne encephalitis virus, rubella virus, and bunyamwera virus (Miller and Krijnse-Locker 2008;Harak and Lohmann 2015;Risco et al 2003;Salanueva et al 2003;Delgui et al 2013;Westerbeck and Machamer 2015), and is frequently fragmented after infection (Campadelli et al 1993;Salanueva et al 2003;Yadav et al 2016;Avitabile et al 1995;Lavi et al 1996;Hansen et al 2017;Rebmann et al 2016). Viruses use Golgi membranes directly and/or hijack master controllers of Golgi biogenesis and trafficking to generate vesicles that are used as the site of viral RNA replication (Quiner and Jackson 2010;Hansen et al 2017;Short et al 2013), wrapping (Sivan et al 2016;Alzhanova and Hruby 2007;Alzhanova and Hruby 2006;Nanbo et al 2018;Lundu et al 2018;Procter et al 2018), intracellular transduction (Nonnenmacher et al 2015), and secretion ).…”

“…Viruses use Golgi membranes directly and/or hijack master controllers of Golgi biogenesis and trafficking to generate vesicles that are used as the site of viral RNA replication (Quiner and Jackson 2010;Hansen et al 2017;Short et al 2013), wrapping (Sivan et al 2016;Alzhanova and Hruby 2007;Alzhanova and Hruby 2006;Nanbo et al 2018;Lundu et al 2018;Procter et al 2018), intracellular transduction (Nonnenmacher et al 2015), and secretion ). Viral infection triggers Golgi fragmentation via diverse mechanisms, ranging from phosphorylating key Golgi structural proteins such as GRASP65 (Rebmann et al 2016), activating the Src kinase to phosphorylate the Dynamin 2 GTPase (Martin et al 2017), targeting the immunity-related GTPase M (IRGM) to the Golgi to induce GBF1 phosphorylation (Hansen et al 2017), modulating vesicular trafficking (Yadav et al 2016;Johns et al 2014), to impeding the major histocompatibility complex (MHC) class I trafficking, antigen presentation, and/or cytokine secretion Rohde et al 2012).…”

Avian infl uenza

“…Step 5,6: Infl uenza viral RNA sections missing a 5 cap for transcription of RNA-dependent RNA polymerase, so the PA ,PB1 and PB2, elements conduct cap-snatching of host DNA to fi nish this process (20-22) Cap-containing viral mRNA is discharged through the host ribosome apparatus into the cytoplasm to bedeciphered. Surface proteins such as NA and HA are changed over into the rough endoplasmic reticulum and translocated for post-translation adjustments into the Golgi apparatus [23].…”