Replicating satellite RNA induces sequence-specific DNA methylation and truncated transcripts in plants
Wang, M.B., Wesley, S.V., Finnegan, E.J., Smith, N.A., & Waterhouse, P.M. (2001) Replicating satellite RNA induces sequence-specific DNA methylation and truncated transcripts in plants. RNA, 7(1), pp. 16-28.
Tobacco plants were transformed with a chimeric transgene comprising sequences encoding β-glucuronidase (GUS) and the satellite RNA (satRNA) of cereal yellow dwarf luteovirus. When transgenic plants were infected with potato leafroll luteovirus (PLRV), which replicated the transgene-derived satRNA to a high level, the satellite sequence of the GUS:Sat transgene became densely methylated. Within the satellite region, all 86 cytosines in the upper strand and 73 of the 75 cytosines in the lower strand were either partially or fully methylated. In contrast, very low levels of DNA methylation were detected in the satellite sequence of the transgene in uninfected plants and in the flanking nonsatellite sequences in both infected and uninfected plants. Substantial amounts of truncated GUS:Sat RNA accumulated in the satRNA-replicating plants, and most of the molecules terminated at nucleotides within the first 60 bp of the satellite sequence. Whereas this RNA truncation was associated with high levels of satRNA replication, it appeared to be independent of the levels of DNA methylation in the satellite sequence, suggesting that it is not caused by methylation. All the sequenced GUS:Sat DNA molecules were hypermethylated in plants with replicating satRNA despite the phloem restriction of the helper PLRV. Also, small, sense and antisense ∼22 nt RNAs, derived from the satRNA, were associated with the replicating satellite. These results suggest that the sequence-specific DNA methylation spread into cells in which no satRNA replication occurred and that this was mediated by the spread of unamplified satRNA and/or its associated 22 nt RNA molecules.
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|Item Type:||Journal Article|
|Additional Information:||Articles free to read on journal website after 12 months|
|Keywords:||Double-stranded RNA, Gene silencing, Luteovirus, RNA interference, RNA-directed DNA methylation, Transcription elongation, cytosine, DNA, elongation factor, satellite RNA, virus RNA, article, DNA methylation, DNA transcription, enzyme activity, gene replication, gene sequence, plant, priority journal, RNA replication, transgene, Base Sequence, Caulimovirus, DNA, Plant, DNA, Satellite, Molecular Sequence Data, Plants, Genetically Modified, Plants, Toxic, Promoter Regions (Genetics), Reverse Transcriptase Polymerase Chain Reaction, Rhizobium, RNA, Plant, RNA, Satellite, Tobacco, Transcription, Genetic, Embryophyta, Nicotiana obtusifolia, Nicotiana tabacum, Potato leafroll virus, Solanum tuberosum, PMCID: PMC1370065|
|Subjects:||Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > PLANT BIOLOGY (060700)|
|Divisions:||Current > Schools > School of Earth, Environmental & Biological Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
|Deposited On:||08 Jan 2014 03:09|
|Last Modified:||29 Jan 2015 05:56|
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