title: The Melolontha melolontha entomopoxvirus (MmEPV) fusolin is related to the fusolins of lepidopteran EPVs and to the 37K baculovirus glycoprotein.
abstract: We have cloned and sequenced a 1.7-kbp DNA fragment of the MmEPV genome encompassing the major polypeptide of the spindle-shaped inclusions gene termed fusolin. The sequence contained a single open reading frame of 1203 nt capable of coding for a polypeptide of 45.8 kDa. The 13 N-terminal amino acid (aa) residues were hydrophobic and could act as a signal peptide. The aa sequence also contained 13 cysteine residues very likely involved in paracrystal formation. This sequence showed significant homologies with the fusolins of two lepidopteran EPVs, the Choristoneura biennis EPV (CbEPV) and the Heliothis armigera EPV, and also with the 37K glycoproteins of Autographa californica and Orgyia pseudotsugata baculoviruses. No homology was found between the MmEPV fusolin and the 100K MmEPV spherulin, nor with the 110K polypeptide of the CbEPV and Amsacta moorei EPV spheroidins. These data were confirmed by Western blot analysis. Transfection of vaccinia-infected mammalian cells with a plasmid encompassing the fusolin sequence plus the upstream regulatory region resulted in transient expression of the gene. This indicated that the vaccinia transcription machinery is able to transcribe the fusolin gene. The fusolin was also expressed in insect cells via a recombinant baculovirus.
authors.affiliation: Proteine Performance S. A., Route d'Alès, Saint Christol lez Alès, France.
authors.forename: L
authors.initials: L
authors.lastname: Gauthier
authors.forename: F
authors.initials: F
authors.lastname: Cousserans
authors.forename: J C
authors.initials: JC
authors.lastname: Veyrunes
authors.forename: M
authors.initials: M
authors.lastname: Bergoin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D017996:Entomopoxvirinae / Q000235:genetics*; D005814:Genes, Viral / Q000235:genetics*; D007181:Inclusion Bodies, Viral / Q000737:chemistry; D007313:Insecta / Q000821:virology*; D007915:Lepidoptera / Q000821:virology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000235:genetics; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: C066445:Ac-gp37 protein, Autographa californica nucleopolyhedrovirus; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C093475:fusolin protein, entomopoxvirus