Transcriptional and translational expression kinetics of the UL25 homologue of bovine herpesvirus 1.1.
We investigated whether the bovine herpesvirus 1.1 (BHV1) ORF, a homologue of the herpes simplex virus 1 (HSV1) UL25 gene, represented a functional gene. The BHV1 UL25 ORF, which is located at positions 60602<--62398 of the viral genome, generated a 4.5 kb transcript accumulating at low abundance as soon as 3 hours p.i. after which the levels increased up to 12h p.i. and remained constant up to 24 hours p.i. In addition, UL25 transcription initiated at 303 bases upstream from the translation initiation codon, corresponding to 26 and 354 b downstream from putative TATA and CAAT boxes, respectively, thus providing evidence that these elements function as the UL25 promoter. Western blotting of BHV1-infected cell lysates, using a BHV1-UL25 specific antiserum generated against a T7-Tag/UL25 fusion recombinant protein expressed in E. coli, detected a 63 kDa protein of the expected size (63,083 Da) whose expression profile followed that of its transcript. The synthesis of the 63 kDa protein was abrogated by a DNA synthesis inhibitor, unambiguously demonstrating that the viral specific protein expressed from the BHV1 UL25 ORF belongs to the gamma2 class.
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Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Québec, Canada.
Boucher
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Simard
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D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002417:Cattle; D002460:Cell Line; D006566:Herpesviridae Infections / Q000821:virology*; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000502:physiology*; D007668:Kidney / Q000166:cytology; D007700:Kinetics; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus