The core 2 β-1,6-N-acetylglucosaminyltransferase-M encoded by bovine herpesvirus 4 is not essential for virus replication despite contributing to post-translational modifications of structural proteins

Markine-Goriaynoff, Nicolas; Gillet, Laurent; Karlsen, Odd André; Haarr, Lars; Minner, Frédéric; Pastoret, Paul-Pierre; Fukuda, Minoru; Vanderplasschen, Alain

doi:10.1099/vir.0.19715-0

Cited by 18 publications

(19 citation statements)

References 46 publications

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“…Subconfluent monolayers of A549 or OVCAR cells were infected with BoHV-4 Vtest strain at a multiplicity of infection (MOI) of 1 pfu per cell. From the time of infection up to the detection of apoptosis, cycloheximide (50 Ag/mL) or phosphonoacetic acid (300 Ag/mL) were added to the culture medium to inhibit de novo protein synthesis or DNA polymerase, respectively (32).…”

Section: Methodsmentioning

confidence: 99%

Bovine Herpesvirus 4 Induces Apoptosis of Human Carcinoma Cell Lines In vitro and In vivo

Gillet

Dewals

Farnir³

et al. 2005

Cancer Research

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The idea of using oncolytic viruses for the treatment of cancers was proposed a century ago. During the last two decades, viruses able to replicate specifically in cancer cells and to induce their lysis were identified and were genetically modified to improve their viro-oncolytic properties. More recently, a new approach consisting of inducing selective apoptosis in cancer cells through viral infection has been proposed; this approach has been called viro-oncoapoptosis. In the present study, we report the property of bovine herpesvirus-4 (BoHV-4) to induce, in vitro and in vivo, apoptosis of some human carcinomas. This conclusion relies on the following observations:

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Section: Methodsmentioning

confidence: 99%

Bovine Herpesvirus 4 Induces Apoptosis of Human Carcinoma Cell Lines In vitro and In vivo

Gillet

Dewals

Farnir³

et al. 2005

Cancer Research

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“…Plaque size. MDBK cells grown on coverslips were infected with BoHV-4 and then overlaid with MEM containing 10% FCS and 0.6% (wt/vol) carboxymethylcellulose (CMC) (medium viscosity) (Sigma) in order to obtain isolated plaques (34). At successive intervals after infection, the plaques were stained by indirect immunofluorescence staining using MAb 35.…”

mentioning

confidence: 99%

“…A BoHV-4 V. test strain deleted for nucleotides 43 to 608 of the Bo10 gene (Bo10 Del) was produced by homologous recombination (34). The restriction fragment EcoRI-B (containing Bo10) of the V. test strain genome was cloned into the pGEM-T Easy vector (Promega), resulting in pGEM-T Easy/EcoRI-B.…”

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confidence: 99%

The Bovine Herpesvirus 4 Bo10 Gene Encodes a Nonessential Viral Envelope Protein That Regulates Viral Tropism through both Positive and Negative Effects

Machiels¹,

Lété²,

Fays³

et al. 2011

J Virol

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All gammaherpesviruses encode a glycoprotein positionally homologous to the Epstein-Barr virus gp350 and the Kaposi's sarcoma-associated herpesvirus (KSHV) K8.1. In this study, we characterized the positional homologous glycoprotein of bovine herpesvirus 4 (BoHV-4), encoded by the Bo10 gene. We identified a 180-kDa gene product, gp180, that was incorporated into the virion envelope. A Bo10 deletion virus was viable but showed a growth deficit associated with reduced binding to epithelial cells. This seemed to reflect an interaction of gp180 with glycosaminoglycans (GAGs), since compared to the wild-type virus, the Bo10 mutant virus was both less infectious for GAG-positive (GAG ؉ ) cells and more infectious for GAG-negative (GAG ؊ ) cells. However, we could not identify a direct interaction between gp180 and GAGs, implying that any direct interaction must be of low affinity. This function of gp180 was very similar to that previously identified for the murid herpesvirus 4 gp150 and also to that of the Epstein-Barr virus gp350 that promotes CD21؉ cell infection and inhibits CD21 ؊ cell infection. We propose that such proteins generally regulate virion attachment both by binding to cells and by covering another receptor-binding protein until they are displaced. Thus, they regulate viral tropism both positively and negatively depending upon the presence or absence of their receptor.Many viruses use a single glycoprotein for both cell binding and membrane fusion. Herpesviruses are more complex. Three proteins-gB, gH, and gL-form a core fusion machinery conserved in the Alpha-, Beta-, and Gammaherpesvirinae subfamilies (21). Most herpesviruses also encode at least one additional receptor-binding protein that is more specific for a given virus subfamily. For example, herpes simplex virus first attaches to cells by gB or gC binding to the heparan sulfate moieties of the cell surface proteoglycans. gD must then bind for fusion to occur (47).Our understanding of gammaherpesvirus glycoprotein functions is more limited. This is due to the fact that the human gammaherpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) show limited lytic growth in vitro. Nevertheless, understanding how viral glycoproteins function is important for vaccination and for the development of neutralizing antibodies. While gB, gH, and gL are probably the key partners of the gammaherpesvirus membrane fusion machinery (22,39,41), the function of accessory entry proteins is less clear. EBV gp350/220 is a highly glycosylated membrane protein that adopts its 2 differently sized forms by alternative splicing (23). It is the most abundant protein in the virion envelope, binds to complement receptor 2 (CD21) on B cells (40,49), and is a target for antibodies that neutralize B cell infection (53). Soluble recombinant gp350 can also inhibit EBV infection of CD21-positive (CD21 ϩ

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“…Then, RT-PCR experiments identified a product of expression corresponding to the full-length coding sequence of the gene and encoding a functional ␤1,6-GnT that formed core 2, core 4, and I structures (13). Expression of this active Bo17 product was confirmed for all available BoHV-4 strains by RT-PCR and a subsequent enzymatic activity assay (18).…”

Section: Resultsmentioning

confidence: 97%

“…The BoHV-4 V.test strain deleted for the Bo17 ORF (Bo17 Del) and the corresponding revertant strain (Bo17 Rev) have been described previously (18). The recombinant strains Bo17 MuDir and Bo17 Spliced were derived from a cloned BoHV-4 bacterial artificial chromosome (BAC) (29).…”

Section: Importancementioning

confidence: 99%

Bovine Herpesvirus 4 Modulates Its β-1,6- N -Acetylglucosaminyltransferase Activity through Alternative Splicing

Lété

Markine-Goriaynoff

Machiels

et al. 2016

J Virol

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Carbohydrates play major roles in host-virus interactions. It is therefore not surprising that, during coevolution with their hosts, viruses have developed sophisticated mechanisms to hijack for their profit different pathways of glycan synthesis. Thus, the Bo17 gene of Bovine herpesvirus 4 (BoHV-4) encodes a homologue of the cellular core 2 protein ␤-1,6-N-acetylglucosaminyltransferase-mucin type (C2GnT-M), which is a key player for the synthesis of complex O-glycans. Surprisingly, we show in this study that, as opposed to what is observed for the cellular enzyme, two different mRNAs are encoded by the Bo17 gene of all available BoHV-4 strains. While the first one corresponds to the entire coding sequence of the Bo17 gene, the second results from the splicing of a 138-bp intron encoding critical residues of the enzyme. Antibodies generated against the Bo17 C terminus showed that the two forms of Bo17 are expressed in BoHV-4 infected cells, but enzymatic assays revealed that the spliced form is not active. In order to reveal the function of these two forms, we then generated recombinant strains expressing only the long or the short form of Bo17. Although we did not highlight replication differences between these strains, glycomic analyses and lectin neutralization assays confirmed that the splicing of the Bo17 gene gives the potential to BoHV-4 to fine-tune the global level of core 2 branching activity in the infected cell. Altogether, these results suggest the existence of new mechanisms to regulate the activity of glycosyltransferases from the Golgi apparatus. IMPORTANCEViruses are masters of adaptation that hijack cellular pathways to allow their growth. Glycans play a central role in many biological processes, and several studies have highlighted mechanisms by which viruses can affect glycosylation. Glycan synthesis is a nontemplate process regulated by the availability of key glycosyltransferases. Interestingly, bovine herpesvirus 4 encodes one such enzyme which is a key enzyme for the synthesis of complex O-glycans. In this study, we show that, in contrast to cellular homologues, this virus has evolved to alternatively express two proteins from this gene. While the first one is enzymatically active, the second results from the alternative splicing of the region encoding the catalytic site of the enzyme. We postulate that this regulatory mechanism could allow the virus to modulate the synthesis of some particular glycans for function at the location and/or the moment of infection.

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The core 2 β-1,6-N-acetylglucosaminyltransferase-M encoded by bovine herpesvirus 4 is not essential for virus replication despite contributing to post-translational modifications of structural proteins

Cited by 18 publications

References 46 publications

Bovine Herpesvirus 4 Induces Apoptosis of Human Carcinoma Cell Lines In vitro and In vivo

Bovine Herpesvirus 4 Induces Apoptosis of Human Carcinoma Cell Lines In vitro and In vivo

The Bovine Herpesvirus 4 Bo10 Gene Encodes a Nonessential Viral Envelope Protein That Regulates Viral Tropism through both Positive and Negative Effects

Bovine Herpesvirus 4 Modulates Its β-1,6- N -Acetylglucosaminyltransferase Activity through Alternative Splicing

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