Time Course Analysis and Mapping of Autographa californica Nuclear Polyhedrosis Virus Transcripts

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Abstract
To study the expression of the Autographa californica nuclear polyhedrosis virus (AcNPV) genome, intracellular virus-specific proteins and mRNA were pulsed-labeled, extracted and analyzed at 6 h intervals during the replicative cycle. Most RNA were detected 12-24 h postinfection (p.i.), but many continued to be synthesized until late in infection. Polyhedrin and p8 mRNA were the 2 most abundant late viral RNA transcripts, and they were synthesized at high rates until late in the infection cycle (60 h p.i.). The abundancy control of polyhedrin and p8 polypeptides was considered to be at the level of transcription. Two other major mRNA in infected Spodoptera frugiperda cells were 0.6-kilobase RNA, which was synthesized at its highest rate 12-18 h p.i., and 2.8-kilobase RNA, which was synthesized from 12-48 h p.i. Cytoplasmic polyA-containing RNA was isolated at 6-h intervals and was analyzed by Northern blot hybridization. At least 50 virus RNA transcripts were recognized, sized and mapped onto the genome. Six RNA hybridized to EcoRI-H, -I and -J, and HindIII-Q AcNPV DNA restriction fragments, 7 RNA hybridized to EcoRI-B and -D DNA fragments, 5 RNA hybridized to EcoRI-A and -E regions of the genome, 4 RNA hybridized to EcoRI-C and -N DNA fragments and 1 RNA species hybridized to EcoRI-O AcNPV DNA. A transcription map of the AcNPV genome was constructed, and the data were correlated with previously published translation maps.

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