Organization of the M genomic segment of Toscana phlebovirus.
The nucleotide sequence of the Toscana (TOS) virus M RNA segment contains a single major open reading frame in the viral-complementary sequence, which can encode a polyprotein of 1339 amino acids. To map the TOS M segment product(s), different regions of the putative M polypeptide were expressed as glutathione S-transferase fusion proteins, which were purified and inoculated into mice to produce hyperimmune sera. By Western blot analysis, a protein of approximately 30 kDa and two glycoproteins, G1 and G2, with the same molecular mass (approximately 65 kDa) were identified in TOS virus-infected cells. The 30 kDa protein, which reacted with antibodies raised to the NH2-terminal, was found to be a non-structural protein (designated NSm). By immunoprecipitation analysis of TOS virus-infected cell lysates, both treated or untreated with tunicamycin, the relative positions of glycoproteins G1 and G2 were determined. The gene order, with respect to the genomic M RNA, was found to be 3' NSm-G1 -G2 5'
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Laboratory of Virology, Istituto Superiore di Sanità, Rome, Italy.
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D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D017931:DNA Primers; D015723:Gene Library; D016679:Genome, Viral; D005982:Glutathione Transferase / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D016856:Phlebovirus / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014709:Vero Cells; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000235:genetics*
D017931:DNA Primers; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D005982:Glutathione Transferase