Identification and analysis of an alcelaphine herpesvirus 1 (AHV-1) cDNA clone expressing a fusion protein recognized by AHV-1-neutralizing antisera

Lahijani, Roya S.; Sutton, Stacy M.; Klieforth, Robert B.; Heuschele, Werner P.

doi:10.1007/bf01718430

Cited by 2 publications

(1 citation statement)

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“…This may also apply to ORF 73, which shares structural similarity to herpesvirus saimiri and HHV-8 ORF 73. A partial cDNA encompassing the 3Ј end of ORF 73 terminates in the 29-bp repetitive region, and a fusion protein derived from this cDNA is recognized by antisera from AHV-1-infected animals (40). The potential membrane protein with homology to herpesvirus saimiri ORF 75 and EBV BNRF1 has highly significant similarities to FGARAT (EC 6.3.5.3) (8), an enzyme catalyzing the fourth step in the de novo synthesis of purine bases.…”

Section: Discussionmentioning

confidence: 99%

Primary structure of the alcelaphine herpesvirus 1 genome

Ensser¹,

Pflanz²,

Fleckenstein³

1997

J Virol

142

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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.

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Section: Discussionmentioning

confidence: 99%