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See detailCloning of the genome of Alcelaphine herpesvirus 1 as an infectious and pathogenic bacterial artificial chromosome.
Dewals, Benjamin G ULg; Boudry, Christel ULg; Gillet, Laurent ULg et al

in Journal of General Virology (The) (2006), 87(Pt 3), 509-17

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order ... [more ▼]

Alcelaphine herpesvirus 1 (AlHV-1), carried asymptomatically by wildebeest, causes malignant catarrhal fever (MCF) following cross-species transmission to a variety of susceptible species of the order Artiodactyla. The study of MCF pathogenesis has been impeded by an inability to produce recombinant virus, mainly due to the fact that AlHV-1 becomes attenuated during passage in culture. In this study, these difficulties were overcome by cloning the entire AlHV-1 genome as a stable, infectious and pathogenic bacterial artificial chromosome (BAC). A modified loxP-flanked BAC cassette was inserted in one of the two large non-coding regions of the AlHV-1 genome. This insertion allowed the production of an AlHV-1 BAC clone stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. The loxP-flanked BAC cassette was excised from the genome of reconstituted virions by growing them in permissive cells stably expressing Cre recombinase. Importantly, BAC-derived AlHV-1 virions replicated comparably to the virulent (low-passage) AlHV-1 parental strain and induced MCF in rabbits that was indistinguishable from that of the virulent parental strain. The availability of the AlHV-1 BAC is an important advance for the study of MCF that will allow the identification of viral genes involved in MCF pathogenesis, as well as the production of attenuated recombinant candidate vaccines. [less ▲]

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See detailEvolution of Bovine herpesvirus 4: recombination and transmission between African buffalo and cattle
Dewals, Benjamin G ULg; Thirion, Muriel ULg; Markine-Goriaynoff, N. et al

in Journal of General Virology (2006), 87(Pt 6), 1509-1519

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world, but virological and serological studies have suggested that the African buffalo is also a natural host for this virus. It ... [more ▼]

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world, but virological and serological studies have suggested that the African buffalo is also a natural host for this virus. It has previously been found that the Bo17 gene of BoHV-4 was acquired from an ancestor of the African buffalo, probably around 1.5 million years ago. Analysis of the variation of the Bo17 gene sequence among BoHV-4 strains suggested a relatively ancient transmission of BoHV-4 from the buffalo to the Bos primigenius lineage, followed by a host-dependent split between zebu and taurine BoHV-4 strains. In the present study, the evolutionary history of BoHV-4 was investigated by analysis of five gene sequences from each of nine strains representative of the viral species: three isolated from African buffalo in Kenya and six from cattle from Europe, North America and India. No two gene sequences had the same evolutionary tree, indicating that recombination has occurred between divergent lineages; six recombination events were delineated for these sequences. Nevertheless, exchange has been infrequent enough that a clonal evolutionary history of the strains could be discerned, upon which the recombination events were superimposed. The dates of divergence among BoHV-4 lineages were estimated from synonymous nucleotide-substitution rates. The inferred evolutionary history suggests that African buffalo were the original natural reservoir of BoHV-4 and that there have been at least three independent transmissions from buffalo to cattle, probably via intermediate hosts and - at least in the case of North American strains - within the last 500 years. [less ▲]

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See detailDevelopment of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning.
Gillet, Laurent ULg; Daix, V.; Donofrio, G. et al

in Journal of General Virology (The) (2005), 86(Pt 4), 907-17

Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been ... [more ▼]

Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been impeded by the difficulty in manipulating its large genome using classical homologous recombination in eukaryotic cells. In the present study, the feasibility of exploiting bacterial artificial chromosome (BAC) cloning and prokaryotic recombination technology for production of BoHV-4 recombinants was explored. Firstly, the BoHV-4 genome was BAC cloned using two potential insertion sites. Both sites of insertion gave rise to BoHV-4 BAC clones stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. Reconstituted virus replicated comparably to wild-type parental virus and the loxP-flanked BAC cassette was excised by growing them on permissive cells stably expressing Cre recombinase. Secondly, BoHV-4 recombinants expressing Ixodes ricinus anti-complement protein I or II (IRAC I/II) were produced using a two-step mutagenesis procedure in Escherichia coli. Both recombinants induced expression of high levels of functional IRAC molecules in the supernatant of infected cells. This study demonstrates that BAC cloning and prokaryotic recombination technology are powerful tools for the development of BoHV-4 as an expression vector and for further fundamental studies of this gammaherpesvirus. [less ▲]

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See detailLes glycosyltransférases virales
Markine-Goriaynoff, N.; Vanderplasschen, Alain ULg

in Virologie (2005), 9(1), 35-48

Les glycanes constituent une classe biochimique difficile à investiguer, mais dont les rôles essentiels à tous les niveaux de la biologie se sont imposés comme une évidence ces dernières décennies. Il ... [more ▼]

Les glycanes constituent une classe biochimique difficile à investiguer, mais dont les rôles essentiels à tous les niveaux de la biologie se sont imposés comme une évidence ces dernières décennies. Il apparaît maintenant qu'ils représentent la quatrième catégorie de molécules bio-informatives après l'ADN, l'ARN et les protéines. À ce titre, après la génomique, la transcriptomique et la protéomique, l'ère de la glycomique est appelée à concentrer beaucoup d'intérêts. Des millions d'années durant, les virus ont co-évolué avec leurs hôtes; au cours de ce processus adaptatif, ils ont évolué de manière à favoriser leur multiplication par l'acquisition de moyens visant à imiter, détourner ou saboter les mécanismes les plus complexes de la physiologie de leurs hôtes, y compris les mécanismes destinés à interférer avec le glycome. La découverte de l'importance du glycome dans le contexte de la régulation des interactions entre les virus et leurs hôtes a récemment mené à la notion de « glycovirologie ». Un aspect fascinant de la glycovirologie est l'étude des mécanismes viraux visant à modifier le glycome. Les virus affectent le glycome de deux façons: en régulant l'expression des glycosyltransférases de la cellule hôte ou en exprimant leur propre glycosyltransférase. Cette revue est consacrée aux glycosyltransférases virales et à la description de leurs fonctions. La description de ces enzymes illustre différents concepts fondamentaux de virologie et démontre indirectement le rôle crucial joué par la glycomique dans les processus biologiques. [less ▲]

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See detailThe core 2 beta-1,6-N-acetylglucosaminyltransferase-mucin encoded by bovine herpesvirus 4 was acquired from an ancestor of the African buffalo
Markine-Goriaynoff, N.; Georgin, J. P.; Goltz, M. et al

in Journal of Virology (2003), 77(3), 1784-1792

The Bo17 gene of bovine herpesvirus 4 (BoHV-4) is the only viral gene known to date that encodes a homologue of the cellular core 2 beta-1,6-N-acetylglucosaminyltransferase-mucin type (C2GnT-M). To ... [more ▼]

The Bo17 gene of bovine herpesvirus 4 (BoHV-4) is the only viral gene known to date that encodes a homologue of the cellular core 2 beta-1,6-N-acetylglucosaminyltransferase-mucin type (C2GnT-M). To investigate the origin and evolution of the Bo17 gene, we analyzed its distribution among BoHV-4 strains and determined the sequences of Bo17 from nine representative strains and of the C2GnT-M gene from six species of ruminants expected to encompass the group within which the gene acquisition occurred. Of 34 strains of BoHV-4, isolated from four different continents, all were found to contain the Bo17 gene. Phylogenetic analyses indicated that Bo17 was acquired from a recent ancestor of the African buffalo, implying that cattle subsequently acquired BoHV-4 by cross-species transmission. The rate of synonymous nucleotide substitution in Bo17 was estimated at 5 x 10(-8) to 6 x 10(-8) substitutions/site/year, consistent with previous estimates made under the assumption that herpesviruses have cospeciated with their hosts. The Bo17 gene acquisition was dated to around 1.5 million years ago. Bo17 sequences from BoHV-4 strains from African buffalo and from cattle formed two separate clades, estimated to have split about 700,000 years ago. Analysis of the ratio of nonsynonymous to synonymous nucleotide substitutions revealed a burst of amino acid replacements subsequent to the transfer of the cellular gene to the viral genome, followed by a return to a strong constraint on nonsynonymous changes during the divergence of contemporary BoHV-4 strains. The Bo17 gene represents the most recent of the known herpesvirus gene acquisitions and provides the best opportunity for learning more about this important process of viral evolution. [less ▲]

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See detailL’herpèsvirus bovin 4
Markine-Goriaynoff, N.; Minner, F.; De Fays, K. et al

in Annales de Médecine Vétérinaire (2003), 147(4), 215-247

Bovine herpesvirus 4 (BoHV-4) belongs to the Herpesviridae family, Gammaherpesvirinae subfamily, Rhadinovirus genus like human herpesvirus 8 (HHV-8), the causative agent of Kaposi's sarcoma. BoHV-4 has a ... [more ▼]

Bovine herpesvirus 4 (BoHV-4) belongs to the Herpesviridae family, Gammaherpesvirinae subfamily, Rhadinovirus genus like human herpesvirus 8 (HHV-8), the causative agent of Kaposi's sarcoma. BoHV-4 has a worldwide distribution in the cattle population. It has been isolated from cattle showing various clinical signs as well as from healthy cattle. The interest of the scientific community for BoHV-4 is explained by two reasons. Firstly, BoHV-4 represents an homologous virus/host species model to study the biology of gammaherpesviruses. Secondly, the use of BoHV-4 as a recombinant vector for expression both in vitro and in vivo has been proposed. For these reasons, a considerable amount of data has been collected on this virus. In the present paper, the authors will present a general overview of the literature published on this virus addressing clinical, epidemiological and fundamental aspects of BoHV-4. Finally, in the light of their recent phylogenetic data, the authors will discuss the origin and the host species of BoHV-4 leading to the conclusion that this virus should be considered as a virus of the African buffalo rather than cattle. [less ▲]

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See detailA Multipotential Beta -1,6-N-Acetylglucosaminyl-Transferase Is Encoded by Bovine Herpesvirus Type 4
Vanderplasschen, Alain ULg; Markine-Goriaynoff, N.; Lomonte, P. et al

in Proceedings of the National Academy of Sciences of the United States of America (2000), 97(11), 5756-5761

The beta-1,6-N-acetylglucosaminyltransferase (beta1,6GnT) gene family encodes enzymes playing crucial roles in glycan synthesis. Important changes in beta1,6GnT expression are observed during development ... [more ▼]

The beta-1,6-N-acetylglucosaminyltransferase (beta1,6GnT) gene family encodes enzymes playing crucial roles in glycan synthesis. Important changes in beta1,6GnT expression are observed during development, oncogenesis, and immunodeficiency. The most characterized beta1,6GnTs in this gene family are the human (h) C2GnT-L and h-IGnT, which have core 2 [Galbeta1-->3(GlcNAcbeta1-->6)GalNAc] and I branching [GlcNAcbeta1-->3(GlcNAcbeta1-->6)Gal] activities, respectively. Recently, h-C2GnT-M was shown to be unique in forming core 2, core 4 [GlcNAcbeta1-->3(GlcNAcbeta1-->6)GalNAc], and I structures. To date, the beta1,6GnT gene family has been characterized only in mammals. Here, we describe that bovine herpesvirus type 4 (BHV-4) encodes a beta1,6GnT expressed during viral replication and exhibiting all of the core 2, core 4, and I branching activities. Sequencing of the BHV-4 genome revealed an ORF, hereafter called BORFF3-4, encoding a protein (pBORFF3-4) exhibiting 81.1%, 50.7%, and 36.6% amino acid identity with h-C2GnT-M, h-C2GnT-L, and h-IGnT, respectively. Reverse transcriptase-PCR analysis revealed that BORFF3-4 is expressed during BHV-4 replication. Expression of BORFF3-4 in Chinese hamster ovary cells directed the expression of core 2 branched oligosaccharides and I antigenic structures on the cell surface. Moreover, a soluble form of pBORFF3-4 had core 4 branching activity in addition to core 2 and I branching activities. Finally, infection of a C2GnT-negative cell line with BHV-4 induced expression of core 2 branched oligosaccharides. This study extends the beta1,6GnT gene family to a viral gene and provides a model to study the biological functions of a beta1,6GnT in the context of viral infection. [less ▲]

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