Abstract
Avocado sunblotch viroid, peach latent mosaic viroid, chrysanthemum chlorotic mottle viroid, and eggplant latent viroid (ELVd), the four recognized members of the family Avsunviroidae, replicate through the symmetric pathway of an RNA-to-RNA rolling-circle mechanism in chloroplasts of infected cells. Viroid oligomeric transcripts of both polarities contain embedded hammerhead ribozymes that, during replication, mediate their self-cleavage to monomeric-length RNAs with 5'-hydroxyl and 2',3'-phosphodiester termini that are subsequently circularized. We report that a recombinant version of the chloroplastic isoform of the tRNA ligase from eggplant (Solanum melongena L.) efficiently catalyzes in vitro circularization of the plus [(+)] and minus [(-)] monomeric linear replication intermediates from the four Avsunviroidae. We also show that while this RNA ligase specifically recognizes the genuine monomeric linear (+) ELVd replication intermediate, it does not do so with five other monomeric linear (+) ELVd RNAs with their ends mapping at different sites along the molecule, despite containing the same 5'-hydroxyl and 2',3'-phosphodiester terminal groups. Moreover, experiments involving transient expression of a dimeric (+) ELVd transcript in Nicotiana benthamiana Domin plants preinoculated with a tobacco rattle virus-derived vector to induce silencing of the plant endogenous tRNA ligase show a significant reduction of ELVd circularization. In contrast, circularization of a viroid replicating in the nucleus occurring through a different pathway is unaffected. Together, these results support the conclusion that the chloroplastic isoform of the plant tRNA ligase is the host enzyme mediating circularization of both (+) and (-) monomeric linear intermediates during replication of the viroids belonging to the family Avsunviroidae.
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Claims 4
Claim · model-written summary
Eggplant chloroplastic tRNA ligase circularized both polarities of Avsunviroidae replication intermediates in vitro.
- Taxon
- eggplant
Solanum melongenae0 - Gene
- tRNA ligasee0
- Outcome
- circularization of the plus [(+)] and minus [(-)] monomeric linear replication intermediatese0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We report that a recombinant version of the chloroplastic isoform of the tRNA ligase from eggplant (Solanum melongena L.) efficiently catalyzes in vitro circularization of the plus [(+)] and minus [(-)] monomeric linear replication intermediates from the four Avsunviroidae.”
Claim · model-written summary
The ligase recognized the genuine ELVd intermediate but not five alternative terminal variants.
- Taxon
- eggplantSolanum melongenae1
- Gene
- RNA ligasee0
- Outcome
- specifically recognizes the genuine monomeric linear (+) ELVd replication intermediatee0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We also show that while this RNA ligase specifically recognizes the genuine monomeric linear (+) ELVd replication intermediate, it does not do so with five other monomeric linear (+) ELVd RNAs with their ends mapping at different sites along the molecule, despite containing the same 5'-hydroxyl and 2',3'-phosphodiester terminal groups.”
e1“tRNA ligase from eggplant (Solanum melongena L.)”
Claim · model-written summary
Silencing plant tRNA ligase reduced ELVd circularization in Nicotiana benthamiana.
- Taxon
- Nicotiana benthamiana Domin
Nicotiana benthamianae0 - Treatment
- silencing of the plant endogenous tRNA ligasee0
- Gene
- tRNA ligasee0
- Outcome
- ELVd circularizatione0
- Direction
- reductione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Moreover, experiments involving transient expression of a dimeric (+) ELVd transcript in Nicotiana benthamiana Domin plants preinoculated with a tobacco rattle virus-derived vector to induce silencing of the plant endogenous tRNA ligase show a significant reduction of ELVd circularization.”
Claim · model-written summary
Circularization of a nuclear-replicating viroid was unaffected.
- Taxon
- Nicotiana benthamiana DominNicotiana benthamianat0
- Outcome
- circularization of a viroid replicating in the nucleuse0
- Direction
- unaffectede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In contrast, circularization of a viroid replicating in the nucleus occurring through a different pathway is unaffected.”
t0“Nicotiana benthamiana Domin plants”
The same questions in other papers
loss of function · organism · Nicotiana benthamianaPositive and negative results
6 papers in the same direction as this paper · 7 in the opposite direction
- negative
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