Regulation of equine herpesvirus type 1 gene expression: characterization of immediate early, early, and late transcription.
The regulation of equine herpesvirus type 1 (EHV-1) transcription was examined in infected rabbit kidney cells using metabolic inhibitors. In order to map EHV-1 immediate early, early, and late transcripts, viral RNA was 32P-labeled in vivo and hybridized to EHV-1 DNA restriction fragments immobilized on nitrocellulose filters. Immediate early viral RNA was mapped to one region of the viral genome within the inverted repeat DNA sequences (map units 0.78-0.83 and 0.95-1.0). Northern blot hybridization analysis using a 32P-labeled cloned DNA probe from this region identified a single immediate early viral transcript (approximately 6 kb). Transcription of early and late genes was not restricted to any specific region on the viral genome as indicated by the ability of 32P-labeled early and late RNA to hybridize to EHV-1 restriction endonuclease fragments from both the long and short components of EHV-1 DNA. Additional experiments performed without the use of metabolic inhibitors confirmed that EHV-1 transcription is temporally regulated. The characterization of EHV-1 transcription during productive infection will serve as a reference for the analysis of viral transcripts in oncogenically transformed and persistently infected cells.
D000818:Animals; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation; D005814:Genes, Viral; D006564:Herpesviridae / Q000235:genetics*; D004861:Herpesvirus 1, Equid / Q000235:genetics* / Q000502:physiology; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000235:genetics; D014158:Transcription, Genetic
D004279:DNA, Viral; D012367:RNA, Viral; D003513:Cycloheximide; D004262:DNA Restriction Enzymes