Transcription analysis of the EcoRI D region of the baculovirus Autographa californica nuclear polyhedrosis virus identifies an early 4-kilobase RNA encoding the essential p143 gene.
We have investigated the transcriptional activity of the 60.1- to 68.3-map-unit region of the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV). Twelve transcripts mapping to this region were expressed at various times during infection. An early 4.0-kb transcript, potentially coding for a 143-kDa peptide essential for viral DNA replication, was maximally abundant at 6 h postinfection (p.i.). Transcripts of 0.5, 1.1, 1.4, 2.1, and 3.1 kb were most abundant at 12 h p.i., while two large transcripts of 5.2 and 6.8 kb were expressed maximally at 24 h p.i. In the presence of cycloheximide, and in ts8-infected cells at the nonpermissive temperature, only the 4.0-kb RNA was expressed. Northern (RNA) blot analysis using DNA subfragments from the EcoRI D fragment as probes suggested that many of the transcripts overlapped. Strand-specific cRNA probes revealed that the majority of the RNAs were transcribed in the counterclockwise direction. S1 nuclease and primer extension analysis were used to map the 5' ends of transcripts coded within the 60.1- to 64.8-map-unit region. Mapping of the 3' ends of the 1.1-, 4.0-, 5.2-, and 6.8-kb transcripts suggested that these RNAs were all coterminal at their 3' ends. A minicistron was found between the early 4.0-kb transcription start site and the predicted ATG start codon of the p143 gene. Several similar sequence motifs were identified in the promoter regions of the p143 gene and the AcMNPV DNA polymerase gene.
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Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
Carstens
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D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D015246:Deoxyribonuclease EcoRI; D005814:Genes, Viral; D007313:Insecta; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016366:Open Reading Frames; D015347:RNA Probes; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D014158:Transcription, Genetic; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication
D009838:Oligodeoxyribonucleotides; D015347:RNA Probes; D012367:RNA, Viral; D015678:Viral Structural Proteins; D015246:Deoxyribonuclease EcoRI