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identification of potyviral amorphous inclusion protein as a nonstructural, virus-specific protein related to helper component.antisera to amorphous inclusion (ai) proteins associated with infections by pepper mottle virus (pemv) and the watermelon mosaic virus-1 strain of papaya ringspot virus (prsv-w) were used to probe in vitro translation products of the viral rnas. the major translation product of pemv rna in the rabbit reticulocyte lysate (rrl) system was a previously reported polypeptide of apparent molecular weight 78,000 (mr 78k). it reacted with anti-ai serum, whereas the major translation product in the wheat ...198518639843
cistron mapping of tobacco vein mottling virus.the location and order of cistrons in the rna of the potyvirus tobacco vein mottling virus (tvmv) were investigated. hybrid-arrested translation, using cloned single-stranded dna probes complementary to various regions of the viral rna, was performed and the resulting translation products were analyzed by electrophoresis. the pattern of polypeptides produced with each probe was different from that of control reactions containing rna alone. immunoprecipitation of reaction products with antisera t ...198618640635
the nucleotide sequence of tobacco vein mottling virus rna.the nucleotide sequence of the rna of tobacco vein mottling virus, a member of the potyvirus group, was determined. the rna was found to be 9471 residues in length, excluding a 3'-terminal poly(a) tail. the first three aug codons from the 5'-terminus were followed by in-frame termination codons. the fourth, at position 206, was the beginning of an open reading frame of 9015 residues which could encode a polyprotein of 340 kda. no other long open reading frames were present in the sequence or its ...19863737407
mapping of the tobacco vein mottling virus vpg cistron.the location of the cistron encoding the genome-linked protein (vpg) in the potyvirus tobacco vein mottling virus (tvmv) was investigated. precipitation of 125i-labeled vpg with anti-tobacco etch virus 49k nuclear inclusion protein antiserum (which reacts with the nia nuclear inclusion protein of tvmv) indicated that the tvmv vpg is immunologically related to nia. lysyl residues were found to be present at positions 2, 11, and 16 of the amino-terminal region of the vpg. a search of the tvmv poly ...19883354210
molecular cloning, sequencing and expression in escherichia coli of the bean yellow mosaic virus coat protein gene.the sequence of 1015 nucleotides from the 3' poly(a) tract of the potyvirus bean yellow mosaic virus (bymv) rna has been determined from two cdna clones. this sequence contained a single long open reading frame (orf) starting upstream of the cloned region. the orf was expressed as a fusion protein in escherichia coli, and the product was detected by antibodies specific for the coat protein of bymv. the predicted length of the coat protein gene was 822 nucleotides, corresponding to a 273 amino ac ...19892671258
the complete nucleotide sequence of plum pox potyvirus rna.the complete nucleotide sequence of the plum pox virus (ppv) rna genome has been determined. the rna sequence is 9786 nucleotides in length, excluding the 3'-terminal poly(a) tail. an aug triplet at position 147-149 was assigned as the initiation codon for the translation of the genome size viral polyprotein which would consist of 3140 amino acid residues. the nucleotide sequence of the non-coding regions and the predicted amino acid sequence of the polyprotein of ppv were compared with those pr ...19892773595
vpg-mediated aggregation of potyviral rna.rna prepared from the potyvirus tobacco vein mottling virus contained aggregates of the 9.5 kb genomic rna with electrophoretic mobilities corresponding to 20 and 41 kb species. similar aggregates were present in preparations of the rnas of two other potyviruses. aggregation occurred during or after purification of the rna by sucrose gradient centrifugation and alcohol precipitation and was dependent upon the presence of a protein apparently bound covalently to a region at or near the 5' terminu ...19911990064
complete nucleotide sequence and genetic organization of papaya ringspot virus rna.the complete nucleotide sequence of the rna genome of papaya ringspot virus (prsv) was determined from four overlapping cdna clones and by direct sequencing of viral rna. the genomic rna is 10326 nucleotides in length, excluding the poly(a) tract, and contains one large open reading frame that starts at nucleotide positions 86 to 88 and ends at positions 10118 to 10120, encoding a polyprotein of 3344 amino acids. the highly conserved sequence aaauaaaanancucaacacaacaua at the 5' end of the rna of ...19921402799
nucleotide sequence of the 3'-terminal region of potato virus a rna.the sequence of the 3'-terminal region of the genome of the potato virus a (pva) was obtained from two independent cdna clones. this sequence is 1383 nucleotides long and contains an open reading frame of 1178 nucleotides, ending with the translation termination codon taa and followed by untranslated region of 205 nucleotides. since the n-terminal amino acid of the coat protein of pva was blocked, the position of the putative coat protein cleavage site has been deduced by searching for consensus ...19921604933
plants that express a potyvirus proteinase gene are resistant to virus infection.transgenic tobacco plants that express the genome-linked protein/proteinase-coding region of the potyvirus tobacco vein mottling virus (tvmv) were produced and tested for their reaction to inoculation with tvmv and two other potyviruses. these plants did not develop disease symptoms after being inoculated with large doses of tvmv but were as susceptible to infection by the other potyviruses as were control plants. lines of tobacco that express the coat protein- or the nonstructural cylindrical i ...19938327491
mutations in the helper component protease gene of zucchini yellow mosaic virus affect its ability to mediate aphid transmissibility.the nucleotide sequence of the helper component protease (hc-pro) genes of three zucchini yellow mosaic virus (zymv) strains has been compared with that of a helper-deficient strain of zymv-hc. the comparisons revealed three unique deduced amino acid differences. two of these mutations were located in regions which are conserved in other potyviruses. the role of these mutations in aphid transmissibility was examined by exchanging dna fragments of part of the deficient hc-pro gene with the respec ...19948207404
molecular cloning, expression, and purification of nuclear inclusion a protease from tobacco vein mottling virus.the gene encoding the c-terminal protease domain of the nuclear inclusion protein a (nia) of tobacco vein mottling virus (tvmv) was cloned from an isolated virus particle and expressed as a fusion protein with glutathione s-transferase in escherichia coli xl1-blue. the 27-kda protease was purified from the fusion protein by glutathione affinity chromatography and mono s chromatography. the purified protease exhibited the specific proteolytic activity towards the nonapeptide substrates, ac-glu-as ...200010850655
comparison of the substrate specificity of two potyvirus proteases.the substrate specificity of the nuclear inclusion protein a (nia) proteolytic enzymes from two potyviruses, the tobacco etch virus (tev) and tobacco vein mottling virus (tvmv), was compared using oligopeptide substrates. mutations were introduced into tev protease in an effort to identify key determinants of substrate specificity. the specificity of the mutant enzymes was assessed by using peptides with complementary substitutions. the crystal structure of tev protease and a homology model of t ...200515654889
molecular cloning, overproduction, purification and biochemical characterization of the p39 nsp2 protease domains encoded by three alphaviruses.alphaviruses cause serious diseases that pose a potential health threat to both humans and livestock. the nonstructural protein 2 (nsp2) encoded by alphaviruses is a multifunctional enzyme that is essential for viral replication and maturation. its 39-kda c-terminal domain (nsp2pro) is a cysteine protease that is responsible for cleaving a viral polyprotein at three sites to generate nonstructural proteins 1, 2, 3 and 4. in the present study, we evaluated nsp2pro domains from the following three ...200919013248
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