title: The pnk/pnl gene (ORF 86) of Autographa californica nucleopolyhedrovirus is a non-essential, immediate early gene.
abstract: Autographa californica nucleopolyhedrovirus (AcMNPV) ORF 86, located within the HindIII C fragment, potentially encodes a protein which shares sequence similarity with two T4 bacteriophage gene products, RNA ligase and polynucleotide kinase. This AcMNPV gene has been designated pnk/pnl but has yet to be assigned a function in virus replication. It has been classified as an immediate early virus gene, since the promoter was active in uninfected insect cells and mRNA transcripts were detectable from 4 to 48 h post-infection and in the presence of cycloheximide or aphidicolin in virus-infected cells. The extremities of the transcript have been mapped by primer extension and 3' RACE-PCR to positions -18 from the translational start codon and +15 downstream of the stop codon. The function of pnk/pnl was investigated by producing a recombinant virus (Acdel86lacZ) with the coding region replaced with that of lacZ. This virus replicated normally in Spodoptera frugiperda (Sf 21) cells, indicating that pnk/pnl is not essential for propagation in these cells. Virus protein production in Acdel86lacZ-infected Sf 21 cells also appeared to be unaffected, with normal synthesis of the IE-1, GP64, VP39 and polyhedrin proteins. Shut-down of host protein synthesis was not abolished in recombinant infection. When other baculovirus genomes were examined for the presence of pnk/pnl by restriction enzyme digestion and PCR, a deletion was found in AcMNPV 1.2, Galleria mellonella NPV (GmMNPV) and Bombyx mori NPV (BmNPV), suggesting that in many isolates this gene has either never been acquired or has been lost during genome evolution. This is one of the first baculovirus immediate early genes that appears to be nonessential for virus survival.
authors.affiliation: Unité de Génétique des Virus, Station de Recherches de Pathologie Comparée, INRA-URA, CNRS 2209, Saint-Christol-les-Alès, France.
authors.forename: D
authors.initials: D
authors.lastname: Durantel
authors.forename: L
authors.initials: L
authors.lastname: Croizier
authors.forename: M D
authors.initials: MD
authors.lastname: Ayres
authors.forename: G
authors.initials: G
authors.lastname: Croizier
authors.forename: R D
authors.initials: RD
authors.lastname: Possee
authors.forename: M
authors.initials: M
authors.lastname: López-Ferber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D017781:Genes, Immediate-Early / Q000235:genetics*; D005814:Genes, Viral; D017874:Immediate-Early Proteins / Q000235:genetics*; D007313:Insecta; D008025:Ligases / Q000235:genetics; D008780:Methyltransferases / Q000378:metabolism; D008969:Molecular Sequence Data; D000077183:Occlusion Body Matrix Proteins; D016133:Polymerase Chain Reaction; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000235:genetics; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera; D014162:Transfection; D014760:Viral Fusion Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D015678:Viral Structural Proteins
chemical_list: C111524:Ac-PNK-PNL protein, Autographa californica nucleopolyhedrovirus; D017874:Immediate-Early Proteins; D000077183:Occlusion Body Matrix Proteins; C088682:VP39 protein, Vaccinia virus; D014760:Viral Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D008780:Methyltransferases; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D008025:Ligases