Abstract
The RNA replicase extracted from Brome mosaic virus (BMV)-infected plants has been used to characterize the cis-acting elements for RNA synthesis and the mechanism of RNA synthesis. Minus-strand RNA synthesis in vitro requires a structure named stem-loop C (SLC) that contains a clamped adenine motif. In vitro, there are several specific requirements for SLC recognition. We examined whether these requirements also apply to BMV replication in barley protoplasts. BMV RNA3s with mutations in SLC were transfected into barley protoplasts, and the requirements for minus- and plus-strand replication were found to correlate well with the requirements in vitro. Furthermore, previous analysis of replicase recognition of the Cucumber mosaic virus (CMV) and BMV SLCs indicates that the requirements in the BMV SLC are highly specific. In protoplasts, we found that BMV RNA3s with their SLCs replaced with two different CMV SLCs were defective for replication. In vitro results generated with the BMV replicase and minimal-length RNAs generally agreed with those of in vivo BMV RNA replication. To extend this conclusion, we determined that, corresponding with the process of infection, the BMV replicases extracted from plants at different times after infection have different levels of recognition of the minimal promoters for plus- and minus-strand RNA syntheses.
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Claims 4
Claim · model-written summary
Mutant BMV RNA3 replication requirements in barley protoplasts correlated with in vitro requirements.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaree0 - Interactor
- BMV
Brome mosaic viruse0pathogen - Gene
- SLCe0
- Outcome
- requirements for minus- and plus-strand replicatione0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“BMV RNA3s with mutations in SLC were transfected into barley protoplasts, and the requirements for minus- and plus-strand replication were found to correlate well with the requirements in vitro”
Claim · model-written summary
Replacing BMV RNA3 SLCs with two CMV SLCs made replication defective in barley protoplasts.
- Taxon
- barleyHordeum vulgaret0
- Treatment
- SLCs replaced with two different CMV SLCse0
- Interactor
- BMVBrome mosaic viruse0pathogen
- Outcome
- defective for replicatione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In protoplasts, we found that BMV RNA3s with their SLCs replaced with two different CMV SLCs were defective for replication”
t0“Barley”
Claim · model-written summary
In vitro minimal RNA results generally agreed with in vivo BMV replication.
- Taxon
- BarleyHordeum vulgaret0
- Interactor
- BMVBrome mosaic viruse0pathogen
- Outcome
- in vitro resultse0
- Comparator
- in vivo BMV RNA replicatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In vitro results generated with the BMV replicase and minimal-length RNAs generally agreed with those of in vivo BMV RNA replication”
t0“Barley”
Claim · model-written summary
BMV replicases extracted at different times after infection differed in recognition of minimal plus- and minus-strand promoters.
- Taxon
- BarleyHordeum vulgaret0
- Interactor
- BMVBrome mosaic viruse0pathogen
- Outcome
- different levels of recognition of the minimal promoters for plus- and minus-strand RNA synthesese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the BMV replicases extracted from plants at different times after infection have different levels of recognition of the minimal promoters for plus- and minus-strand RNA syntheses”
t0“Barley”