Identification of novel transcripts complementary to the Marek's disease virus homologue of the ICP4 gene of herpes simplex virus.
Libraries of cDNA were generated from polyadenylated RNAs derived from Marek's disease virus (MDV)-transformed cell lines by directional cloning of oligo-(dT)-primed cDNAs in lambda gt22A. Analysis of the libraries for viral sequences showed that a number of cDNA clones originated from transcripts mapping in the BamHI A region of the MDV genome. Sequencing and fine mapping of these cDNAs suggested that the RNA transcripts expressed from this region were either in the sense or antisense direction with respect to the MDV homologue of the ICP4 gene of herpes simplex virus. The longest cDNA clone from antisense transcripts was 2756 bp long and partially overlapped the 5' end of the coding region of the ICP4 gene. The cDNA clone contained at least four introns, shown by comparison of its sequence with the sequence of the ICP4 gene. The presence of introns was confirmed by PCR analysis. All the introns have the consensus splice donor and acceptor signals at their 5' and 3' ends respectively. Northern blot analysis showed that the ICP4 gene homologue of MDV was abundantly transcribed only in lytically infected fibroblasts, whereas transcripts complementary to the ICP4 gene were the major transcripts in MDV-transformed cell lines and lymphomas. The transcripts complementary to ICP4 consist of two major RNA species approximately 15 kb and 1.32 kb long. The results suggest that there might be an inverse relationship between the abundance of ICP4 transcripts and their complementary transcripts in MDV-infected and transformed cells.
AFRC Institute for Animal Health, Compton Laboratory, Newbury, Berkshire, U.K.
D S
DS
Li
J
J
Pastorek
V
V
Zelník
G D
GD
Smith
L J
LJ
Ross
D000595:Amino Acid Sequence; D001483:Base Sequence; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary; D015723:Gene Library; D016679:Genome, Viral; D008381:Herpesvirus 2, Gallid / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000235:genetics; D007438:Introns; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D018139:Simplexvirus / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014764:Viral Proteins
D018076:DNA, Complementary; D017874:Immediate-Early Proteins; D009687:Nuclear Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D012367:RNA, Viral; D015534:Trans-Activators; D014764:Viral Proteins; C048925:herpes simplex virus, type 1 protein ICP4; C075882:ICP4 protein, Gallid herpesvirus 2