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Galiakparov, N., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Goszczynski, D.E., Virology Unit, Agricultural Research Council, Plant Protection Research Institute, Pretoria, South Africa
Che, X., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Batuman, O., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Bar-Joseph, M., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Mawassi, M., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Grapevine virus A (GVA), a species of the recently established genus Vitivirus, consists of an ∼7.3-kb single-stranded RNA genome of positive polarity, organized into five open reading frames (ORFs). The virus, which is closely associated with the grapevine rugose wood disease complex, has been poorly investigated genetically. We explored the production of viral RNAs in a GVA-infected Nicotiana benthamiana herbaceous host and characterized one nested set of three 5′-terminal sgRNAs of 5.1, 5.5, and 6.0 kb, and another, of three 3′-terminal sgRNAs of 2.2, 1.8, and 1.0 kb that could serve for expression of ORFs 2-3, respectively. Neither 3′- nor 5′-terminal sgRNAs, which would correspond to ORF5, was detected, suggesting that expression of this ORF occurs via a bi- or polycistronic mRNA. The 5′-terminal sgRNAs were abundant in dsRNA-enriched extracts. Cloning and sequence analysis of the 3′ end of 5.5-kb 5′-terminal sgRNA and the 5′ end of the 1.8-kb 3′-terminal sgRNA suggested that a mechanism other than specific cleavage was involved in production of these sgRNAs. Apparently, the production of the 5′- and 3′-terminal sgRNAs was controlled by sequences upstream of the 5′-terminus of each of ORFs 2-4. Detection of both plus and minus strands of the 5′- and 3′-terminal sgRNAs, though in different levels of accumulation, suggested that each of these cis-acting elements is involved in production of four RNAs: a 3′-terminal plus-strand sgRNA which could act as an mRNA, the corresponding 3′-terminal minus-strand RNA, a 5′-terminal plus-strand sgRNA, and the corresponding 5′-terminal minus-strand RNA. © 2003 Elsevier Science (USA). All rights reserved.
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Two classes of subgenomic RNA of grapevine virus A produced by internal controller elements
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Galiakparov, N., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Goszczynski, D.E., Virology Unit, Agricultural Research Council, Plant Protection Research Institute, Pretoria, South Africa
Che, X., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Batuman, O., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Bar-Joseph, M., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Mawassi, M., Department of Virology, Agricultural Research Organization, Volcani Center, 50250 Bet Dagan, Israel
Two classes of subgenomic RNA of grapevine virus A produced by internal controller elements
Grapevine virus A (GVA), a species of the recently established genus Vitivirus, consists of an ∼7.3-kb single-stranded RNA genome of positive polarity, organized into five open reading frames (ORFs). The virus, which is closely associated with the grapevine rugose wood disease complex, has been poorly investigated genetically. We explored the production of viral RNAs in a GVA-infected Nicotiana benthamiana herbaceous host and characterized one nested set of three 5′-terminal sgRNAs of 5.1, 5.5, and 6.0 kb, and another, of three 3′-terminal sgRNAs of 2.2, 1.8, and 1.0 kb that could serve for expression of ORFs 2-3, respectively. Neither 3′- nor 5′-terminal sgRNAs, which would correspond to ORF5, was detected, suggesting that expression of this ORF occurs via a bi- or polycistronic mRNA. The 5′-terminal sgRNAs were abundant in dsRNA-enriched extracts. Cloning and sequence analysis of the 3′ end of 5.5-kb 5′-terminal sgRNA and the 5′ end of the 1.8-kb 3′-terminal sgRNA suggested that a mechanism other than specific cleavage was involved in production of these sgRNAs. Apparently, the production of the 5′- and 3′-terminal sgRNAs was controlled by sequences upstream of the 5′-terminus of each of ORFs 2-4. Detection of both plus and minus strands of the 5′- and 3′-terminal sgRNAs, though in different levels of accumulation, suggested that each of these cis-acting elements is involved in production of four RNAs: a 3′-terminal plus-strand sgRNA which could act as an mRNA, the corresponding 3′-terminal minus-strand RNA, a 5′-terminal plus-strand sgRNA, and the corresponding 5′-terminal minus-strand RNA. © 2003 Elsevier Science (USA). All rights reserved.
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