Expression and functional analysis of a baculovirus gene encoding a truncated protein kinase homolog.
Autographa californica nuclear polyhedrosis virus (AcMNPV) potentially encodes a 215-amino acid polypeptide containing 6 out of 11 motifs conserved among eukaryotic protein kinases (Morris et al., Virology 200, 360-369, 1994). We examined the expression of this gene, named pk2, at the transcriptional and translational levels and the possible role of the gene during baculovirus replication in cell culture and insect larvae. Northern (RNA) blot analysis revealed that pk2 was transcribed primarily as an early 1.2-kb RNA. Western blot analysis showed that pk2 was expressed as a 25-kDa protein, PK2, which was present both early and late during virus infection. To examine the function(s) of pk2, we constructed a mutant baculovirus, vKINdel, in which one-third of the PK2-coding region was deleted and then compared the characteristics of vKINdel with wild-type AcMNPV and a marker-rescued revertant. The pk2 deletion mutation had no discernable effect on the number, size, or appearance of plaques, the kinetics of protein synthesis or protein phosphorylation profiles during virus infection of cultured SF-21 cells. Deletion of pk2 also had no significant influence on the infectivity or virulence of the baculovirus in larval bioassays and the level of occluded virus production was normal. Thus, pk2 does not appear to have a significant influence on virus replication in the host systems examined.
Li
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Department of Genetics, University of Georgia, Athens 30602.
Miller
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LK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers; D005814:Genes, Viral; D007814:Larva / Q000201:enzymology; D008969:Molecular Sequence Data; D009154:Mutation; D017924:Nucleopolyhedroviruses / Q000201:enzymology / Q000235:genetics*; D010766:Phosphorylation; D011494:Protein Kinases / Q000096:biosynthesis / Q000235:genetics*; D017384:Sequence Deletion; D018411:Spodoptera; D014779:Virus Replication / Q000235:genetics
D017931:DNA Primers; D011494:Protein Kinases