Primary structure of the alcelaphine herpesvirus 1 genome

Abstract

Alcelaphine herpesvirus 1 (AHV-1) causes wildebeest-associated malignant catarrhal fever, a lymphoproliferative syndrome in ungulate species other than the natural host. Based on biological properties and limited structural data, it has been classified as a member of the genus Rhadinovirus of the subfamily Gammaherpesvirinae. Here, we report on cloning and structural analysis of the complete genome of AHV-1 C500. The low GC content DNA (L-DNA) region of the genome consists of 130,608 bp with low (46.17%) GC content and marked suppression of CpG dinucleotide frequency. Like in herpesvirus saimiri, the prototype of the rhadinoviruses, the L-DNA is flanked by approximately 20 to 25 GC-rich (71.83%) high GC content DNA (H-DNA) repeats of 1,113 to 1,118 nucleotides. The analysis of the L-DNA sequence revealed 70 open reading frames (ORFs), 61 of which showed homology to other herpesviruses. The conserved ORFs are arranged in four blocks collinear to other Rhadinovirus genomes. These gene blocks are flanked by nonconserved regions containing ORFs without similarities to known herpesvirus genes. Notably, a spliced reading frame with a coding capacity for a 199-amino-acid protein is located in a position homologous to the transforming genes of herpesvirus saimiri at the left end of the L-DNA. A gene with homology to the semaphorin family is located adjacent to this. Despite common biological and epidemiological properties, AHV-1 differs significantly from herpesvirus saimiri with regard to cell homologous genes, probably using a different set of effector proteins to achieve a similar T-lymphocyte-transforming phenotype. MATERIALS AND METHODS Cell lines and viral culture. The bovine epithelial kidney cell line MDBK (ATCC CCL 22) was propagated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 100 g of gentamicin per ml, and 350 g of L-glutamine per ml. AHV-1 strains C500 (62) and WC11 (63) were serially propagated by infection of fresh MDBK cells with aliquots of infected MDBK cells showing cytopathic changes in more than 80% of the cells. AHV-1 C500 was obtained from H. W. Reid (Moredun Research Institute, Edinburgh, United Kingdom), and the attenuated WC11 strain was obtained from D. W. Verwoerd (Veterinary Research Institute, Onderstepoort, South Africa). For the preparation of virions, tissue culture supernatant was precleared by centrifugation at 2,000 ϫ g. Strain C500 virions (passage 5) were pelleted by centrifugation at 50,000 ϫ g with an SW28 rotor. The viral DNA was extracted from the pellet with phenol-chloroform and precipitated with ethanol. WC11 DNA was further purified by density gradient centrifugation as described previously (37). Cloning procedures.

Molecular and Histopathological Characterization of Sheep-Associated Malignant Catarrhal Fever (SA-MCF) Outbreak in Beef Cattle

“…These viruses are the causative agents of both wildebeest (or African)‐associated MCF and sheep‐associated MCF, respectively (Plowright et al., ; McGeoch et al., ). A complete genome sequence of both viruses (AlHV‐1, OvHV‐2) has been published, with the genomes consisting of 130 608 and 135 621 bp, respectively (Ensser et al., ; Taus et al., ; Meier‐Trummer et al., ).…”

Prevalence of Bovine Herpesvirus-4 Infection in Cats in Central Michigan

“…41 Wildebeest-associated malignant catarrhal fever is a fatal lymphoproliferative disease of cattle and other large ruminants caused by the gamma-2 herpesvirus Alcelaphine herpesvirus 1. 42 The fact that BHV-4 shares many genetic and biologic features with other gamma-2 herpesviruses raises the question of whether BHV-4 has a potential role in neoplastic or lymphoproliferative diseases. Additional historical, physical examination, clinicopathological, and postmortem examination findings would be of value in investigating the pathogenic role of BHV-4 in cats.…”

Seroprevalence of malignant catarrhal fever virus in captive wildebeest (Connochaetessp.)in France

“…Total DNA was extracted from cryopreserved PBMC cell suspensions using the QIAamp miniprep kit (Qiagen). DNA samples were then subjected to a two‐step nested PCR to amplify a sequence in the ORF50 of the AlHV‐1 genome with primers C500‐1 and C500‐2 for the first reaction, and C500‐3 and C500‐4 for the second reaction (Ensser, Pflanz, & Fleckenstein, ; Li, Dyer, Keller, & Crawford, ). In brief, 1 μg total DNA was used in a total of 50 μl of amplification reaction of Phusion High‐Fidelity DNA polymerase master mix (New England Biolabs) containing 0.4 μM of primer (Eurogentec) and 1 μl DMSO.…”