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Genome sequencing of alcelaphine herpesvirus 1 C500 BAC clone and WC11 attenuated strain identified duplicated and deleted regions involving ORF50, A6, A7 and A8
Myster, Françoise; van Beurden, Steven; Davison, Andrew et al.
2015Fourth annual meeting of the Belgian Society for Virology
 

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Abstract :
[en] Alcelaphine herpesvirus 1 (AlHV-1) is a gammaherpesvirus carried asymptomatically by wildebeest. When it is cross-transmitted to other ruminants, they develop a lymphoproliferative disease named malignant catarrhal fever (MCF). Though the expression of the latency-associated protein encoded by ORF73 has been shown to be essential for MCF induction, the pathogenic mechanisms inducing the lymphoproliferation are yet to be identified. The AlHV-1 genome consists in a long unique region (LUR) containing 73 ORFs flanked by polyrepetitive DNA units (prDNA). AlHV-1 can be cultured in vitro and rearrangements of the AlHV-1 genome have been described after multiple passages in cell culture and are associated with increased cell-free viral particles and loss of pathogenicity. In this study, we sequenced the genomes of the infectious and pathogenic AlHV-1 C500 BAC clone as well as the high-passage attenuated WC11 strain. First, the full sequence of the AlHV-1 BAC clone was obtained using 454 deep sequencing. As expected, the obtained sequence for the AlHV-1 C500 BAC strain was nearly identical to the sequence of the C500 reference strain. However, we identified the translocation of a duplicated region containing ORF50, A6 gene and part of A7 gene flanked by prDNA units. Restriction profile, southern blotting and PCR analyses showed that the duplicated region was separated from the left end of the LUR by one unit of prDNA. We used the BAC clone to remove the translocated region in order to determine whether the duplication of ORF50 and A6 could increase viral fitness in vitro and/or in vivo. Secondly, the full sequence of the WC11 strain was obtained using an Illumina sequencing approach. Although the genome of the WC11 strain was nearly identical to the reference C500 sequence, two regions were deleted. First, a 1,175 bp sequence consists in the left end of the LUR including the predicted A1 gene. Second, a 2,174 bp sequence includes the full coding sequence of A7 gene and the first part of A8 gene. A high sequence divergence was also observed in the right-end of the LUR, a region including the A9.5 and A10 coding sequences. Interestingly, a translocation of a duplicated region containing ORF50 and A6 was also present in the WC11 genome. The genes modified or absent in the WC11 strain are particularly susceptible to be involved in the induction of the disease. While the duplication of ORF50 could have been selected in vitro to enhance viral growth, the potential function of the A1, A9.5, and A10 genes are unknown. However, A7 and A8 genes are positional homolog to the Epstein-Barr virus genes encoding gp42 and gp350. As such, A7 and A8 are potentially involved in the entry of the virus and in the regulation of the viral tropism. Viruses impaired for the expression of A7 and/or A8 are currently in production in order to study their functions in MCF induction.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Myster, Françoise ;  Université de Liège - ULiège > Département des maladies infectieuses et parasitaires (DMI) > Vaccinologie vétérinaire
van Beurden, Steven
Davison, Andrew
Vanderplasschen, Alain ;  Université de Liège - ULiège > Immunologie et vaccinologie
Dewals, Benjamin G  ;  Université de Liège - ULiège > Immunologie et vaccinologie
Language :
English
Title :
Genome sequencing of alcelaphine herpesvirus 1 C500 BAC clone and WC11 attenuated strain identified duplicated and deleted regions involving ORF50, A6, A7 and A8
Publication date :
December 2015
Event name :
Fourth annual meeting of the Belgian Society for Virology
Event date :
December 2015
Available on ORBi :
since 13 January 2020

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