Immediate-early, early, and late RNAs in bovine herpesvirus-4-infected cells

We have begun to identify immediate-early (IE), early (E), and late (L) genes of BHV-4 by analysis of cytoplasmic polyadenylated RNA transcribed from the BHV-4 genome over the course of infection and in the presence of cycloheximide or phosphonoacetic acid. Labeled cDNA prepared from RNA isolated at different times was hybridized with Southern blots of viral DNA to show which regions of the genome are transcribed at different times. In a second series of experiments, radiolabeled cloned restriction fragments representing the entire BHV-4 genome were hybridized separately to RNA on Northern blots to determine the number and sizes of transcripts at different times. As expected, with increasing time after infection, more portions of the BHV-4 genome are transcribed and a larger number and a greater abundance of viral transcripts are present. RNA transcribed from terminal repeats was not detectable at any time. However, similarity in size of RNA transcribed from opposite ends of the unique region of the genome late in infection suggests that RNA is transcribed over the fused ends of the genome, and terminal repeats are removed during RNA processing. Identification of IE, E, and L transcripts by this analysis lays the foundation for further study of specific BHV-4 transcripts and genes.