Origin and Messenger Activity of Two Small RNA Species Found in Particles of Tobacco Rattle Virus Strain SYM

Robinson, David J.; Mayo, M. A.; Fritsch, C.; Jones, A. T.; Raschké, J. H.

doi:10.1099/0022-1317-64-7-1591

Cited by 27 publications

(20 citation statements)

References 20 publications

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“…Short (S) particles of I6, N5, TRV, PEBV-SHE and PEBV-D were obtained by two cycles of centrifugation in sucrose gradients containing 17 mM-phosphate pH 7.8, as described by Kurppa et al (1981). RNA was extracted from virus particles as described by Robinson et al (1983), and from leaf tissue of N. clevelandii infected with NM-type isolates as described by Harrison & Robinson (1982).…”

Section: Methodsmentioning

confidence: 99%

Two Anomalous Tobravirus Isolates: Evidence For RNA Recombination in Nature

Robinson

Hamilton²,

Harrison

et al. 1987

Journal of General Virology

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100

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SUMMARY16 and N5 are naturally occurring tobravirus isolates that produce symptoms in herbaceous plants similar to those induced by strains of tobacco rattle virus (TRV). In immunosorbent electron microscopy tests, however, they reacted with antisera to particles of pea early-browning virus (PEBV), not TRV. Furthermore, these tests indicated that 16 was related to the British serotype of PEBV and N5 to the Dutch. Pseudo-recombinant isolates were produced by reassortment of the genome parts of 16 or N5 with those of TRV, in any combination, but not in most combinations with those of PEBV. However, I6 RNA-2 was replicated in plants inoculated also with RNA-1 from an isolate of the British serotype of PEBV, but the PEBV RNA-1 was imperfectly packaged by 16 coat protein, and the virus particles seemed to have only limited stability. Nucleic acid hybridization experiments showed that the RNA-1 sequences of both I6 and N5 were similar to those of TRV strains. I6 RNA-2 contained sequences resembling those of the British serotype of PEBV, but with some TRV-like sequences at the 3' and 5' ends, whereas N5 RNA-2 contained more extensive TRV-like 3' and 5' ends flanking sequences that were related, but perhaps not closely, to those of the Dutch serotype of PEBV. Thus, the RNA-2 species of 16 and N5 were recombinant molecules that contained sequences typical of both TRV and PEBV, and which probably had separate but similar evolutionary origins. As a result of their hybrid nature, I6 and N5 were part of the gene pool and had the pathogenicity of TRV, while possessing the serological properties of PEBV.

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Section: Methodsmentioning

confidence: 99%

Two Anomalous Tobravirus Isolates: Evidence For RNA Recombination in Nature

Robinson

Hamilton²,

Harrison

et al. 1987

Journal of General Virology

Self Cite

100

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“…After 60 min incubation at 30 °C translation products were analysed in 8 to 20 % gradient polyacrylamide gels using the Tris-glycine-SDS system of Laemmli (1970). Immunoprecipitation of 35S-labelled translation products with the anti-PSLV serum was done as described by Robinson et al (1983).…”

Section: Methodsmentioning

confidence: 99%

Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome

Agranovsky

Karasev

Novikov

et al. 1992

Journal of General Virology

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RNA from the Hungarian isolate of poa semilatent virus (PSLV) directed in vitro synthesis of 120K, 75K, 25K (coat protein) and 20K polypeptides. In vitro translation of PSLV RNA was blocked by the cap analogue, m7Gpp, thus suggesting that the virus RNA was capped. PSLV RNA could be aminoacylated with [~4C]tyrosine in vitro. The sequence of 1-5 kb from the 3' end of the PSLV RNA y component revealed two open reading frames (ORFs) separated by a uridinerich intergenic region. The putative product of the incomplete 5'-proximal ORF showed a close amino acid sequence similarity with the C-terminal segment of the 7a protein (putative RNA replicase) encoded in the barley stripe mosaic virus (BSMV) RNA 7, and the 20K product of the 3'-proximal ORF was found to be related to the 17K 7b product of BSMV. The sequence of 0-8 kb from the 3' end ofPSLV RNA fl encompassed two (incomplete) overlapping ORFs whose putative products are related to the tic and fld proteins encoded in the similarly arranged ORFs of BSMV RNA ft. Nucleotide sequence homology between the respective parts of the two hordeivirus genomes was restricted to the ORF for 7a, the spacer between the ORFs for ya and yb, and the 3' non-coding region, particularly the 95 nucleotide segment at the 3' end representing a tRNAlike structure. Despite limited sequence conservation beyond this segment, the entire 3' non-coding region of PSLV RNA could be folded in a tight pseudoknotted structure closely resembling that of BSMV RNA. Surprisingly, the 'signature' sequence typical for BSMV RNA, internal polydisperse poly(A) intercalated between the coding part and the 3' tRNA-like structure, was not detected in the PSLV genome. Instead, the virus RNA contained several oligoadenylate stretches spaced by other residues, close to the junction of its coding and 3' non-coding portions.

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“…Translation products were analysed in 8 to 20~ polyacrylamide gradient slab gels according to Laemmli (1970) and visualized by autoradiography. Immunoprecipitation of 35S-labelled products was done essentially as described by Robinson et al (1983).…”

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confidence: 99%