Glycoprotein B of equine herpesvirus type 1 has two recognition sites for subtilisin-like proteases that are cleaved by furin

Spiesschaert, Bart; Stephanowitz, Heike; Krause, Eberhard; Osterrieder, Nikolaus; Azab, Walid

doi:10.1099/jgv.0.000418

Cited by 4 publications

(4 citation statements)

References 52 publications

(52 reference statements)

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“…Across the members of the Herpesviridae , gB possesses a conserved cleavage site Arg-X-Lys/Arg-Arg targeted by the cellular protease furin. Here, we show that EEHV-1 gB has also a furin cleavage site like other members [17]. gB of EEHV-1A has a conserved furin cleavage motif 433 RRKR 436 , which cleaves gB in the middle.…”

Section: Discussionmentioning

confidence: 94%

“…Sample buffer (1 M Tris-HC1 (pH 6.8), 0.8% SDS, 0.4% glycerol, 0.15% β-mercaptoethanol, 0.004% bromophenol blue) was added to protein lysates. The samples were heated at 95 °C for 10 min and proteins were separated by 12% SDS-PAGE as described previously [17]. Expression of gB was detected with anti-gB antibodies, which either recognize the peptide sequence located between amino acids 259 and 274 (Ab7125) or between 427 and 441 (Ab7123) of EEHV-1 gB as derived from its sequence (GenBank accession NO.…”

Section: Methodsmentioning

confidence: 99%

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Fatal Elephant Endotheliotropic Herpesvirus Infection of Two Young Asian Elephants

Pavulraj

Eschke

Prahl³

et al. 2019

Microorganisms

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Elephant endotheliotropic herpesvirus (EEHV) can cause a devastating haemorrhagic disease in young Asian elephants worldwide. Here, we report the death of two young Asian elephants after suffering from acute haemorrhagic disease due to EEHV-1A infection. We detected widespread distribution of EEHV-1A in various organs and tissues of the infected elephants. Enveloped viral particles accumulated within and around cytoplasmic electron-dense bodies in hepatic endothelial cells were detected. Attempts to isolate the virus on different cell cultures showed limited virus replication; however, late viral protein expression was detected in infected cells. We further showed that glycoprotein B (gB) of EEHV-1A possesses a conserved cleavage site Arg-X-Lys/Arg-Arg that is targeted by the cellular protease furin, similar to other members of the Herpesviridae. We have determined the complete 180 kb genome sequence of EEHV-1A isolated from the liver by next-generation sequencing and de novo assembly. As virus isolation in vitro has been unsuccessful and limited information is available regarding the function of viral proteins, we have attempted to take the initial steps in the development of suitable cell culture system and virus characterization. In addition, the complete genome sequence of an EEHV-1A in Europe will facilitate future studies on the epidemiology and diagnosis of EEHV infection in elephants.

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Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Fatal Elephant Endotheliotropic Herpesvirus Infection of Two Young Asian Elephants

Pavulraj

Eschke

Prahl³

et al. 2019

Microorganisms

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“…PBMCs were infected with either parental (EHV-1 or EHV-4) or mutant (EHV-1∆US3) viruses (MOI = 1) or transfected with different expression vectors (pcDNA3, pcDNA3-US3_1 or pcDNA3-US3_4; transfection efficiency was around 40%) using Nucleofector™ (Lonza, Köln, Germany) [30]. Twenty-four hours after infection or transfection, cells were incubated with anti-VLA-4 or anti-LFA-1 antibodies (5 μg/mL; Abcam, Cambridge, UK) for 1 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

Proft

Spiesschaert

Izume

et al. 2016

Viruses

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The serine-threonine protein kinase encoded by US3 gene (pUS3) of alphaherpesviruses was shown to modulate actin reorganization, cell-to-cell spread, and virus egress in a number of virus species. However, the role of the US3 orthologues of equine herpesvirus type 1 and 4 (EHV-1 and EHV-4) has not yet been studied. Here, we show that US3 is not essential for virus replication in vitro. However, growth rates and plaque diameters of a US3-deleted EHV-1 and a mutant in which the catalytic active site was destroyed were significantly reduced when compared with parental and revertant viruses or a virus in which EHV-1 US3 was replaced with the corresponding EHV-4 gene. The reduced plaque sizes were consistent with accumulation of primarily enveloped virions in the perinuclear space of the US3-negative EHV-1, a phenotype that was also rescued by the EHV-4 orthologue. Furthermore, actin stress fiber disassembly was significantly more pronounced in cells infected with parental EHV-1, revertant, or the recombinant EHV-1 expressing EHV-4 US3. Finally, we observed that deletion of US3 in EHV-1 did not affect the expression of adhesion molecules on the surface of infected cells.

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“…Most herpes viruses have at least one PC cleavage site motif in the cleavage loop, although, HHSV1 is an exception by having no PC motifs at all. In contrast, other viruses have more than one PC cleavage site, which may be cleaved sequentially [35]. The experimental inactivation of the PC cleavage site of several herpes viruses did not severely affect viral cell entry into cells growing in vitro; however, the lack of PC cleavage reduced virus spread and replication in vivo [36,37].…”

Section: Herpesvirusesmentioning

confidence: 99%

The Proteolytic Regulation of Virus Cell Entry by Furin and Other Proprotein Convertases

Izaguirre

2019

Viruses

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144

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A wide variety of viruses exploit furin and other proprotein convertases (PCs) of the constitutive protein secretion pathway in order to regulate their cell entry mechanism and infectivity. Surface proteins of enveloped, as well as non-enveloped, viruses become processed by these proteases intracellularly during morphogenesis or extracellularly after egress and during entry in order to produce mature virions activated for infection. Although viruses also take advantage of other proteases, it is when some viruses become reactive with PCs that they may develop high pathogenicity. Besides reacting with furin, some viruses may also react with the PCs of the other specificity group constituted by PC4/PC5/PACE4/PC7. The targeting of PCs for inhibition may result in a useful strategy to treat infections with some highly pathogenic viruses. A wide variety of PC inhibitors have been developed and tested for their antiviral activity in cell-based assays.

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Glycoprotein B of equine herpesvirus type 1 has two recognition sites for subtilisin-like proteases that are cleaved by furin

Cited by 4 publications

References 52 publications

Fatal Elephant Endotheliotropic Herpesvirus Infection of Two Young Asian Elephants

Fatal Elephant Endotheliotropic Herpesvirus Infection of Two Young Asian Elephants

The Role of the Equine Herpesvirus Type 1 (EHV-1) US3-Encoded Protein Kinase in Actin Reorganization and Nuclear Egress

The Proteolytic Regulation of Virus Cell Entry by Furin and Other Proprotein Convertases

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