Two novel subgenomic RNAs derived from RNA 3 of tomato aspermy cucumovirus.

Shi, Bu‐Jun; Ding, Shou‐Wei; Symons, Robert H.

doi:10.1099/0022-1317-78-3-505

Cited by 28 publications

(20 citation statements)

References 43 publications

(44 reference statements)

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“…This hotspot is exactly at the 59-end of the conserved motif Box-1 (Blanchard et al, 1996;Thompson et al, 2008), which is common to all subgroup II CMV strains and to TAV but is absent in subgroup I CMV strains (Palukaitis et al, 1992;Shi et al, 1997), and plays a key role in the production of subgenomic RNA5 (de Wispelaere & Rao, 2009;Thompson et al, 2008). Moreover, it has been reported as a hotspot for viral recombination between CMV RNA3 and TAV RNAs 1 and 2 (Suzuki et al, 2003) or TAV RNA3 (de Wispelaere et al, 2005), between TAV RNA3 and CMV RNAs 1 and 2 (Shi et al, 2007), and in Q-CMV (subgroup II) intragenomic recombinants (de Wispelaere & Rao, 2009).…”

Section: Discussion Effects Of 6 Nt Deletions In I17f-cmv Rna3mentioning

confidence: 99%

Analysis of recombination between viral RNAs and transgene mRNA under conditions of high selection pressure in favour of recombinants

Morroni

Thompson

Tepfer

2009

Journal of General Virology

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One possible environmental risk related to the utilization of virus-resistant transgenic plants expressing viral sequences is the emergence of new viruses generated by recombination between the viral transgene mRNA and the RNA of an infecting virus. This hypothesis has been tested recently for cucumber mosaic virus (CMV) by comparing the recombinant populations in transgenic and non-transgenic plants under conditions of minimal selection pressure in favour of the recombinants. Equivalent populations were observed in transgenic and non-transgenic plants but, in both, there was a strongly dominant hotspot recombinant which was shown recently to be nonviable alone in planta, suggesting that its predominance could be reduced by applying an increased selection pressure in favour of viable recombinants. Partially disabled I17F-CMV mutants were created by engineering 6 nt deletions in five sites in the RNA3 39-non-coding region (39-NCR). One mutant was used to inoculate transgenic tobacco plants expressing the coat protein and 39-NCR of R-CMV. A total of 22 different recombinant types were identified, of which 12 were, as expected, between the transgene mRNA and the mutated I17F-CMV RNA3, while 10 resulted from recombination between the mutated RNA3 and I17F-CMV RNA1. Twenty recombinants were of the aberrant type, while two, including the dominant one detected previously under conditions of minimal selection pressure, were homologous recombinants. All recombinants detected were very similar to ones observed in nature, suggesting that the deployment of transgenic lines similar to the one studied here would not lead to the emergence of new viruses.

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Section: Discussion Effects Of 6 Nt Deletions In I17f-cmv Rna3mentioning

confidence: 99%

Analysis of recombination between viral RNAs and transgene mRNA under conditions of high selection pressure in favour of recombinants

Morroni

Thompson

Tepfer

2009

Journal of General Virology

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“…It is interesting to consider whether the TLSs of other viruses are involved in capsid formation. Significantly, short subgenomic RNAs about 300 nt long from the 3Ј ends of CMV and tomato aspermy virus RNAs are very efficiently copackaged into virions with genomic RNAs (33,34), perhaps reflecting a TLS involvement in packaging in this sister genus to the bromoviruses.…”

Section: Discussionmentioning

confidence: 99%

tRNA elements mediate the assembly of an icosahedral RNA virus

Choi

Dreher

Rao

2002

Proc. Natl. Acad. Sci. U.S.A.

100

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tRNAs, the adapter molecules in protein synthesis, also serve as metabolic cofactors and as primers for viral RNA-directed DNA synthesis. The genomic and subgenomic RNAs of some plant viruses have a 3-terminal tRNA-like structure (TLS) that can accept a specific amino acid and serve as a site for initiation of replication and as a simple telomere. We report a previously undescribed role for the TLS of brome mosaic virus (BMV), and potentially for cellular tRNA, in mediating the assembly of its icosahedral virions. BMV genomic RNAs and subgenomic RNA lacking the TLS failed to assemble into virions when incubated with purified BMV coat protein. Assembly was restored by addition of a 201-nt RNA containing the BMV TLS. TLSs from two other plant viruses as well as tRNAs from wheat germ and yeast were similarly active in the BMV virion assembly reaction, but ribosomal RNA and polyadenylate did not facilitate assembly. Surprisingly, virions assembled from TLS-less BMV RNA in the presence of tRNAs or TLS-containing short RNA did not incorporate the latter molecules. Consistent with a critical role for the BMV TLS in virion assembly, mutations in the BMV genomic RNAs that were designed to disrupt the folding of the TLS also abolished virion assembly. We discuss the likely roles of the TLS in early stages of virion assembly.tRNA-like structure ͉ bromoviruses ͉ virus assembly ͉ RNA packaging T ransfer RNAs (tRNAs) are multifunctional. Their primary role is to be an adapter molecule that translates the codon sequences in mRNA into the amino acid sequence of a protein. tRNAs also participate in specialized functions in cellular metabolism such as biosynthesis of the bacterial cell wall (1), chlorophyll, and heme (2). tRNAs and tRNA-related activities are associated with a variety of RNA viruses, including several plant viruses and the retroviruses (3, 4). Host tRNAs found in the virions of retroviruses function as primers for RNA-directed DNA synthesis (3). The tRNA-like structures (TLSs) found at the 3Ј end of the genomes of some plant viruses serve as efficient origins of replication and as primitive telomeres to ensure that the 3Ј-terminal CCA nucleotides are not lost during replication (4, 5). It has been suggested that these viral TLSs are molecular fossils that may relate to a primordial role for tRNA in RNA replication in the ancient RNA world (6).Brome mosaic virus (BMV) is an example of an RNA virus that has an Ϸ170-nt-long TLS covalently bound to the 3Ј end of its genomic and subgenomic RNAs. BMV is a member of the plant virus family Bromoviridae (7) and the alphavirus-like superfamily of human-, animal-, and plant-infecting positive-strand RNA viruses (8). Mature BMV virions encapsidate three genomic RNAs (B1-B3) and a single subgenomic RNA, B4 (7). Physical and biochemical data suggest that B1-B4 are packaged into three morphologically indistinguishable virus particles: B1 (3.2 kb) and B2 (2.9 kb) are packaged individually into separate particles, whereas the genomic B3 (2.1 kb) and the subgenomic B4 (0.9 kb) are...

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“…The probe used to detect the genomic and subgenomic RNAs of TAV was a 32 Plabeled transcript complementary to nucleotides 2287-2386 of RNA 3 of V-TAV [6]. Direct RNA sequencing was performed with primers BJ20, BJ28 and BJ22 (Table 1) according to Fichot and Girard [7].…”

Section: Plant Inoculation and Progeny Viral Rna Analysesmentioning

confidence: 99%

“…Total RNAs were extracted from each infected plant at two weeks after inoculation and were analyzed by Northern blot hybridization as described [6]. The probe used to detect the genomic and subgenomic RNAs of TAV was a 32 Plabeled transcript complementary to nucleotides 2287-2386 of RNA 3 of V-TAV [6].…”

Section: Plant Inoculation and Progeny Viral Rna Analysesmentioning

confidence: 99%

The Conserved, 5? Termini of RNAs 1 and 2 of Tomato aspermy virus are Dispensable for Infection but Affect Virulence

Shi

Palukaitis²,

Symons

2005

Virus Genes

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The 5' terminus of each of the three genomic RNAs (RNAs 1, 2 and 3) of Tomato aspermy virus (TAV) begins with the sequence 5'-GUUU, which is also shared by a number of other viruses. Mutagenic analyses showed that the 5'-GUUU sequence of RNAs 1 and 2 of TAV was dispensable for viral infection and did not prevent symptom induction. On the other hand, substitution of U at position 5 for G in RNA 1, but not RNA 2, induced veinal necrosis symptoms in Nicotiana glutinosa. The mutants constructed included insertion of UUU into the 5'-GUUU sequence of TAV RNAs 1 and/or 2. All RNA 2 mutants induced more severe symptoms than viral RNAs containing either mutated RNA 1 or most combinations of mutated RNAs 1 and 2. Some combinations of mutated RNAs 1 and 2 also induced veinal necrosis in N. glutinosa. Virulence was unrelated to the levels of viral RNA accumulation. Sequence analysis of progeny viral RNAs showed that only the mutant viral RNAs with a G to U substitution in RNA 1 and the deletion of the 5'-GUUU in both RNAs 1 and 2 were able to maintain the same sequence as the inoculum. The other mutants either reverted to the wildtype sequence or underwent further deletion or insertion. None of the constructed mutants were able to compete for accumulation with the wildtype virus after co-inoculation to the plant species tested.

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Two novel subgenomic RNAs derived from RNA 3 of tomato aspermy cucumovirus.

Cited by 28 publications

References 43 publications

Analysis of recombination between viral RNAs and transgene mRNA under conditions of high selection pressure in favour of recombinants

Analysis of recombination between viral RNAs and transgene mRNA under conditions of high selection pressure in favour of recombinants

tRNA elements mediate the assembly of an icosahedral RNA virus

The Conserved, 5? Termini of RNAs 1 and 2 of Tomato aspermy virus are Dispensable for Infection but Affect Virulence

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