A Viral Protein that Blocks Arf1-Mediated COP-I Assembly by Inhibiting the Guanine Nucleotide Exchange Factor GBF1

Wessels, Els; Duijsings, Daniël; Niu, Ting-Kuang; Neumann, Steffi; Oorschot, Viola; Lange, Frank de; Lanke, Kjerstin; Klumperman, Judith; Henke, Andreas; Jackson, Catherine; Melchers, Willem J. G.; Kuppeveld, Frank J. M. van

doi:10.1016/j.devcel.2006.06.005

Cited by 147 publications

(249 citation statements)

References 46 publications

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“…To this end, we co-transfected cells with plasmids encoding FAPP1-PH-GFP together with wt or mutant 3A-myc, and one day later treated them for 1 h with the compounds. In untreated cells, wt 3A was visible in a punctate pattern ( Figure 6A), corresponding to a localization to dispersed Golgi membranes, as previously described [20,23]. 3A showed a great overlap with PI4P, as detected by FAPP1-PH-GFP.…”

Section: Mutations In 3a Do Not Rescue the Activity Of Pi4kiiiβ In Thmentioning

confidence: 67%

“…As primary antibodies we used rabbit polyclonal anti-PI4-KinaseIIIβ (Upstate), mouse monoclonal anti-GBF1 (BD Biosciences), rabbit polyclonal anti-Arf1 (kind gift from dr. F. Wieland, Biochemie-Zentrum Heidelberg, Germany), mouse monoclonal anti-c-Myc (Sigma Aldrich), mouse monoclonal anti-dsRNA (J2, English & Scientific Consulting), mouse anti-PI4P IgM (Echelon Biosciences), mouse monoclonal anti-HA (Covance), mouse monoclonal anti-β-actin (Sigma), and rabbit polyclonal anti-CVB3 3A described previously [23]. The secondary antibodies for immunofluorescence included Alexa Fluor 488-or 594-conjugated goatanti-rabbit IgG, Alexa Fluor 488-or 594-conjugated goat-antimouse IgG, or Alexa Fluor 594-conjugated goat-anti-mouse IgM (Molecular Probes).…”

Section: Antibodiesmentioning

confidence: 99%

“…When expressed in isolation, 3A disrupts ER-to-Golgi transport by blocking the formation of COP-I coat complexes on membranes [18][19][20][21]. The 3A protein most likely exerts this effect by directly interacting with GBF1 [22,23].…”

Section: Introductionmentioning

confidence: 99%

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Coxsackievirus mutants that can bypass host factor PI4KIIIβ and the need for high levels of PI4P lipids for replication

et al. 2012

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RNA viruses can rapidly mutate and acquire resistance to drugs that directly target viral enzymes, which poses serious problems in a clinical context. Therefore, there is a growing interest in the development of antiviral drugs that target host factors critical for viral replication, since they are unlikely to mutate in response to therapy. We recently demonstrated that phosphatidylinositol-4-kinase IIIβ (PI4KIIIβ) and its product phosphatidylinositol-4-phosphate (PI4P) are essential for replication of enteroviruses, a group of medically important RNA viruses including poliovirus (PV), coxsackievirus, rhinovirus, and enterovirus 71. Here, we show that enviroxime and GW5074 decreased PI4P levels at the Golgi complex by directly inhibiting PI4KIIIβ. Coxsackievirus mutants resistant to these inhibitors harbor single point mutations in the non-structural protein 3A. These 3A mutations did not confer compound-resistance by restoring the activity of PI4KIIIβ in the presence of the compounds. Instead, replication of the mutant viruses no longer depended on PI4KIIIβ, since their replication was insensitive to siRNA-mediated depletion of PI4KIIIβ. The mutant viruses also did not rely on other isoforms of PI4K. Consistently, no high level of PI4P could be detected at the replication sites induced by the mutant viruses in the presence of the compounds. Collectively, these findings indicate that through specific single point mutations in 3A, CVB3 can bypass an essential host factor and lipid for its propagation, which is a new example of RNA viruses acquiring resistance against antiviral compounds, even when they directly target host factors.

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Section: Mutations In 3a Do Not Rescue the Activity Of Pi4kiiiβ In Thmentioning

confidence: 67%

Section: Antibodiesmentioning

confidence: 99%

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Coxsackievirus mutants that can bypass host factor PI4KIIIβ and the need for high levels of PI4P lipids for replication

et al. 2012

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“…ARF1, GBF1, and COPI during their successful replication [6,16]. The overexpression of Arf1 or GBF1 reversed the effects of coxsackie virus 3A protein by not inhibiting the trafficking in between ER-Golgi [33]. Yamayoshi et al [37] also showed that dominant-negative mutants of the GTPase Rab1b inhibited the Ebola virus matrix protein VP40-mediated particle formation, consistent the use of this vesicular pathway in the exit of this filovirus.…”

Section: Discussionmentioning

confidence: 95%

Influenza infection modulates vesicular trafficking and induces Golgi complex disruption

et al. 2016

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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.

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“…For viral protein synthesis, recruitment of ribosome to the internal ribosomal entry site (IRES) on the viral genome requires host factors polypyrimidine tract-binding protein (PTB), Unr, La, poly(rC)-binding protein 2 (PCBP2) and SRp20 that are not required for cap-dependent protein synthesis of cellular mRNA (Bedard et al, 2007;Hellen et al, 1993;Hunt et al, 1999;Meerovitch et al, 1993;Sanford et al, 2004;Walter et al, 1999), along with proteolytic modification of a translation initiation factor eIF4G by viral proteinase 2A pro to hijack the translation machinery from cap-dependent protein synthesis to viral protein synthesis (Gradi et al, 1998;Krausslich et al, 1987). For the replication process, membrane rearrangement utilizing ADP ribosylation factors (Arfs) and guanine nucleotide exchange factors (GEFs) (GBF1, BIG1/2) are required via direct and indirect interaction with viral 3A and 3CD proteins in coxsackievirus B3 (CVB3) and PV infections (Belov et al, 2007;Wessels et al, 2006). Cellular signalling via extracellular signal-regulated kinase (ERK), phosphatidylinositol 3-kinase (PI3K) and stress-activated protein kinases (SAPKs) affected replication and/or the release of progeny virus in CVB3 infection by an unknown mechanism (Esfandiarei et al, 2004;Luo et al, 2002;Si et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Characterization of pharmacologically active compounds that inhibit poliovirus and enterovirus 71 infectivity

Arita

Wakita

Shimizu

2008

Journal of General Virology

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Poliovirus (PV) and enterovirus 71 (EV71) cause severe neurological symptoms in their infections of the central nervous system. To identify compounds with anti-PV and anti-EV71 activities that would not allow the emergence of resistant mutants, we performed drug screening by utilizing a pharmacologically active compound library targeting cellular factors with PV and EV71 pseudoviruses that encapsidated luciferase-encoding replicons. We have found that metrifudil (N-[2-methylphenyl]methyl)-adenosine) (an A2 adenosine receptor agonist), N 6 -benzyladenosine (an A1 adenosine receptor agonist) and NF449 (4,49,40,409-[carbonylbis[imino-5,1,3-benzenetriyl bis(carbonyl-imino)]] tetrakis (benzene-1,3-disulfonic acid) octasodium salt) (a Gs-a inhibitor) have anti-EV71 activity, and that GW5074 (3-(3, 5-dibromo-4-hydroxybenzylidine-5-iodo-1,3-dihydro-indol-2-one)) (a Raf-1 inhibitor) has both anti-PV and anti-EV71 activities. EV71 mutants resistant to metrifudil, N 6 -benzyladenosine and NF449 were isolated after passages in the presence of these compounds, but mutants resistant to GW5074 were not isolated for both PV and EV71. The inhibitory effect of GW5074 was not observed in Sendai virus infection and the treatment did not induce the expression of OAS1 and STAT1 mRNA. Small interfering RNA treatment against putative cellular targets of GW5074, including Raf-1, B-Raf, Pim-1, -2, and -3, HIPK2, GAK, MST2 and ATF-3, did not consistently suppress PV replication. Moreover, downregulation of Raf-1 and B-Raf did not affect the sensitivity of RD cells to the inhibitory effect of GW5074. These results suggest that GW5074 has strong and selective inhibitory effect against the replication of PV and EV71 by inhibiting conserved targets in the infection independently of the interferon response.

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A Viral Protein that Blocks Arf1-Mediated COP-I Assembly by Inhibiting the Guanine Nucleotide Exchange Factor GBF1

Cited by 147 publications

References 46 publications

Coxsackievirus mutants that can bypass host factor PI4KIIIβ and the need for high levels of PI4P lipids for replication

Coxsackievirus mutants that can bypass host factor PI4KIIIβ and the need for high levels of PI4P lipids for replication

Influenza infection modulates vesicular trafficking and induces Golgi complex disruption

Characterization of pharmacologically active compounds that inhibit poliovirus and enterovirus 71 infectivity

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