Cloning and sequencing of the spherulin gene, the occlusion body major polypeptide of the Melolontha melolontha entomopoxvirus (MmEPV).
In the late stage of infection, virions of the Melolontha melolontha entomopoxvirus (MmEPV) are occluded into cytoplasmic paracrysalline proteinaceous occlusion bodies designated spherules (A. Amargier, C. Vago, G. Meynadier, 1964, Mikroskopie 19, 309-315). We have cloned and sequenced a 4-kpb DNA fragment of the MmEPV genome encompassing the spherule major protein gene named spherulin. The spherulin gene contains an open reading frame able to code for a 942-amino-acid (aa) polypeptide (MW 109 kDa), consistent with a size above 100 kDa determined by SDS-PAGE for purified spherulin. The MmEPV spherulin showed more than 40% as homology with the Amsacta moorei EPV (AmEPV) spheroidin and shared homologies with the partially sequenced Choristoneura biennis EPV (CbEPV) spheroidin, indicating that this biologically important polypeptide is well conserved among EPVs infecting phylogenetically as distant groups of insects as lepidoptera and coleoptera. Western blot analyses confirmed the relationships between the three polypeptides. In contrast, no homology was detected between the MmEPV spherulin and EPV fusolins or vertebrate poxvirus A-type inclusion proteins. The 45 bases upstream from the ATG initiation codon of spherulin shared 60% homology with the vaccinia virus late promoters including the highly conserved TAAATG consensus sequence. Furthermore, the 5' extremity of the spherulin mRNA consisted of a poly(A) tract of about 20 nucleotides just upstream from the AUG translational initiation codon. These are characteristic features of vertebrate poxvirus late mRNAs suggesting similar modalities of gene expression for vertebrate and insect poxvirus genomes.
Sanz
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Station de Recherche de Pathologie Comparée, INRA-UA CNRS 1184, Saint Christol-Lez-Alés, France.
Veyrunes
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Cousserans
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Bergoin
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003046:Coccidioidin / Q000235:genetics*; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D017996:Entomopoxvirinae / Q000235:genetics*; D005814:Genes, Viral; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral; C009303:spherulin; D003046:Coccidioidin