title: A novel spliced gene in alcelaphine herpesvirus 1 encodes a glycoprotein which is secreted in vitro.
abstract: Herpesviruses often contain cryptic, spliced genes that are not obvious from the initial in silico annotation. Alcelaphine herpesvirus 1 (AlHV-1) contains 72 annotated ORFs but there are also a number of gaps between these that may have protein-coding potential. Comparative analysis of coding potential between AlHV-1 and the related ovine herpesvirus 2 (OvHV-2) revealed a putative novel spliced gene that we have termed A9.5. Analysis of cDNA clones from AlHV-1-infected cells revealed three overlapping clones corresponding to A9.5 and the coding sequence was confirmed by reverse transcription PCR of RNA from AlHV-1-infected cattle tissues. The A9.5 gene was predicted to encode a secreted glycoprotein with molecular mass 19 kDa. Empirical analysis showed that a recombinant haemagglutinin-tagged A9.5 fusion protein was secreted from transfected cells and had a molecular mass of 45 kDa, which was reduced to 20 kDa by endoglycosidase F treatment, confirming that A9.5 was a secreted glycoprotein. In situ RNA hybridization showed that A9.5 was expressed in cells associated with malignant catarrhal fever (MCF) lesions in infected cattle. Detailed analysis of the available OvHV-2 sequences revealed an homologous gene (Ov9.5) with conserved splicing signals and predicted amino acid sequence features in both sequenced isolates of this related virus. We have therefore identified a novel spliced gene in two related macaviruses that is expressed in MCF lesions. Future work will determine its importance for the pathogenesis of disease.
authors: Moredun Research Institute, Pentlands Science Park, Penicuik EH26 0PZ, UK.
authors: George C
authors: GC
authors: Russell
authors: Moredun Research Institute, Pentlands Science Park, Penicuik EH26 0PZ, UK.
authors: Helen
authors: H
authors: Todd
authors: Moredun Research Institute, Pentlands Science Park, Penicuik EH26 0PZ, UK.
authors: David
authors: D
authors: Deane
authors: Moredun Research Institute, Pentlands Science Park, Penicuik EH26 0PZ, UK.
authors: Ann
authors: A
authors: Percival
authors: Moredun Research Institute, Pentlands Science Park, Penicuik EH26 0PZ, UK.
authors: Mark P
authors: MP
authors: Dagleish
authors: School of Veterinary Medicine and Science, Nottingham University, Sutton Bonington, Leicestershire LE12 5RD, UK.
authors: David M
authors: DM
authors: Haig
authors: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool Science Park IC2, Liverpool L3 5RF, UK.
authors: James P
authors: JP
authors: Stewart
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D004274:DNA, Recombinant / Q000235:genetics*; D018142:Gammaherpesvirinae / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006566:Herpesviridae Infections / Q000662:veterinary / Q000821:virology; D007668:Kidney / Q000821:virology; D008304:Malignant Catarrh / Q000821:virology*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
chemical_list: D004274:DNA, Recombinant; D006023:Glycoproteins