Interactions of the Vaccinia Virus A19 Protein

Satheshkumar, Panayampalli S.; Olano, L. Renee; Hammer, Carl H.; Zhao, Min; Moss, Bernard

doi:10.1128/jvi.01261-13

Cited by 10 publications

(17 citation statements)

References 45 publications

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“…Although any part of the duplicated region in RifR9 could be responsible for resistance to rifampin, we focused on A17 because previous studies had shown an interaction between D13 and the N terminus of A17 (5,14), whereas A18 is a DNA-dependent helicase involved in transcription (35,36) and A19 is involved in a late stage of morphogenesis (37,38). A17 is a 23-kDa protein that undergoes several modifications, including C-and N-terminal cleavages and phosphorylation.…”

Section: Isolation Of a Gene Duplication Mutant With Rifampin Resistamentioning

confidence: 99%

Duplication of the A17L Locus of Vaccinia Virus Provides an Alternate Route to Rifampin Resistance

Erlandson

Cotter

Charity

et al. 2014

J Virol

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Specific gene duplications can enable double-stranded DNA viruses to adapt rapidly to environmental pressures despite the low mutation rate of their high-fidelity DNA polymerases. We report on the rapid positive selection of a novel vaccinia virus genomic duplication mutant in the presence of the assembly inhibitor rifampin. Until now, all known rifampin-resistant vaccinia virus isolates have contained missense mutations in the D13L gene, which encodes a capsid-like scaffold protein required for stabilizing membrane curvature during the early stage of virion assembly. Here we describe a second pathway to rifampin resistance involving A17, a membrane protein that binds and anchors D13 to the immature virion. After one round of selection, a rifampin-resistant virus that contained a genomic duplication in the A17L-A21L region was recovered. The mutant had both C-terminally truncated and full-length A17L open reading frames. Expression of the truncated A17 protein was retained when the virus was passaged in the presence of rifampin but was lost in the absence of the drug, suggesting that the duplication decreased general fitness. Both forms of A17 were bound to the virion membrane and associated with D13. Moreover, insertion of an additional truncated or inducible full-length A17L open reading frame into the genome of the wild-type virus was sufficient to confer rifampin resistance. In summary, this report contains the first evidence of an alternate mechanism for resistance of poxviruses to rifampin, indicates a direct relationship between A17 levels and the resistance phenotype, and provides further evidence of the ability of double-stranded DNA viruses to acquire drug resistance through gene duplication. IMPORTANCEThe present study provides the first evidence of a new mechanism of resistance of a poxvirus to the antiviral drug rifampin. In addition, it affirms the importance of the interaction between the D13 scaffold protein and the A17 membrane protein for assembly of virus particles. Resistance to rifampin was linked to a partial duplication of the gene encoding the A17 protein, similar to the resistance to hydroxyurea enabled by duplication of the gene encoding the small subunit of ribonucleotide reductase and of the K3L gene to allow adaptation to the antiviral action of protein kinase R. Gene duplication may provide a way for poxviruses and other DNA viruses with high-fidelity DNA polymerases to adjust rapidly to changes in the environment. Poxviruses are large, enveloped, double-stranded DNA viruses that replicate within the cytoplasm (1). Vaccinia virus (VACV), the model poxvirus, is being employed to construct recombinant vaccines, study antiviral mechanisms, and investigate the basic biology of poxviruses, including assembly. Assembly of VACV begins as viral membranes form and curve into a crescent shape, which is stabilized by a honeycomb lattice of capsid-like D13 protein trimers (2-4, 40). The crescents enlarge to form spheres, termed immature virions (IVs) that, upon processing of the A17 mem...

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Section: Isolation Of a Gene Duplication Mutant With Rifampin Resistamentioning

confidence: 99%

Duplication of the A17L Locus of Vaccinia Virus Provides an Alternate Route to Rifampin Resistance

Erlandson

Cotter

Charity

et al. 2014

J Virol

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“…The two CxxC motifs involved in binding of the VACV orthologue to viral RNA polymerase 29,30 are well conserved in FPV184. Although not investigated in this study, the presence of CxxC motifs is indicative of a zinc finger motif 32,33 .…”

Section: Discussionmentioning

confidence: 99%

“…FPV184 is a well-conserved orthologue of VACV A19 (~ 40% amino acid identity, Supplementary Fig. S5), an essential, late structural protein found in vertebrate but not insect poxviruses 29,30 . To ask if VACV A19 could substitute for FPV184, VACV A19 was overexpressed in human HEK293T cells infected with FWPV∆184 ( Fig.…”

Section: Ectopic Expression Of Fpv184 and Its Ortholog Vacv A19 Resmentioning

confidence: 99%

Modulation of early host innate immune response by a Fowlpox virus (FWPV) lateral body protein

Giotis

Laidlaw

Bidgood

et al. 2020

Preprint

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The avian pathogen, fowlpox virus (FWPV) has been successfully used as vaccine vector in poultry and humans but relatively little is known about its ability to modulate host antiviral immune responses in these hosts, which are replication permissive and non-permissive, respectively. FWPV is highly resistant to avian type I interferon (IFN) and able to completely block the host IFN-response. Microarray screening of host IFN-regulated gene expression in cells infected with 59 different, non-essential FWPV gene knock-out mutants revealed that FPV184 confers immunomodulatory capacity. We report that FPV184-knockout virus (FWPVΔ184) induces the cellular IFN response as early as 2 hours post-infection. The wild-type, uninduced phenotype can be rescued by transient expression of FPV184 in FWPVΔ184-infected cells. Ectopic expression of FPV184 inhibited polyI:C activation of the chicken IFN-β promoter and IFN-α activation of the chicken Mx promoter. Confocal and correlative super-resolution light and electron microscopy demonstrated that FPV184 has a functional nuclear localisation signal domain and is packaged in the lateral bodies of the virions. Taken together, these results provide a paradigm for a late poxvirus structural protein packaged in the lateral bodies and capable of supressing IFN induction early during the next round of infection.

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“…Tryptic peptides were extracted from the gel plugs with formic acid for 30 min at 37°C, followed by lyophilization in SUN-SRI microsampling vials with a SpeedVac. Nano-liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was performed as previously described [ 31 ]. Briefly, lyophilized samples were resuspended in solvent A (0.1% formic acid, 2% AcCN), and peptides were separated on a ProXeon Easy-nLC I multidimensional liquid chromatograph with a linear gradient of solvent A and solvent B (0.1% formic acid, 97.9% AcCN).…”

Section: Methodsmentioning

confidence: 99%

Identification of Outer Membrane and Exoproteins of Carbapenem-Resistant Multilocus Sequence Type 258 Klebsiella pneumoniae

et al. 2015

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Carbapenem-resistant Klebsiella pneumoniae strains have emerged as a cause of life-threatening infections in susceptible individuals (e.g., transplant recipients and critically ill patients). Strains classified as multilocus sequence type (ST) 258 are among the most prominent causes of carbapenem-resistant K. pneumoniae infections worldwide, but the basis for the success of this lineage remains incompletely determined. To gain a more comprehensive view of the molecules potentially involved in the success of ST258, we used a proteomics approach to identify surface-associated and culture supernatant proteins produced by ST258. Protein samples were prepared from varied culture conditions in vitro, and were analyzed by a combination of two-dimensional electrophoresis and liquid chromatography followed by tandem mass spectrometry (LC-MS/MS). We identified a total of 193 proteins in outer membrane preparations from bacteria cultured in Luria-Bertani broth (LB) or RPMI 1640 tissue culture media (RPMI). Compared with LB, several iron-acquisition proteins, including IutA, HmuR, HmuS, CirA, FepA, FitA, FoxA, FhuD, and YfeX, were more highly expressed in RPMI. Of the 177 proteins identified in spent media, only the fimbrial subunit, MrkA, was predicted to be extracellular, a finding that suggests few proteins (or a limited quantity) are freely secreted by ST258. Notably, we discovered 203 proteins not reported in previous K. pneumoniae proteome studies. In silico modeling of proteins with unknown function revealed several proteins with beta-barrel transmembrane structures typical of porins, as well as possible host-interacting proteins. Taken together, these findings contribute several new targets for the mechanistic study of drug-resistance and pathogenesis by ST258 K. pneumoniae isolates.

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Interactions of the Vaccinia Virus A19 Protein

Cited by 10 publications

References 45 publications

Duplication of the A17L Locus of Vaccinia Virus Provides an Alternate Route to Rifampin Resistance

Duplication of the A17L Locus of Vaccinia Virus Provides an Alternate Route to Rifampin Resistance

Modulation of early host innate immune response by a Fowlpox virus (FWPV) lateral body protein

Identification of Outer Membrane and Exoproteins of Carbapenem-Resistant Multilocus Sequence Type 258 Klebsiella pneumoniae

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