Hairpin RNAs derived from RNA polymerase II and polymerase III promoter-directed transgenes are processed differently in plants
Wang, M.B., Helliwell, C.A., Wu, L.M., Waterhouse, P.M., Peacock, W.J., & Dennis, E.S. (2008) Hairpin RNAs derived from RNA polymerase II and polymerase III promoter-directed transgenes are processed differently in plants. RNA, 14(5), pp. 903-913.
RNA polymerase III (Pol III) as well as Pol II (35S) promoters are able to drive hairpin RNA (hpRNA) expression and induce target gene silencing in plants. siRNAs of 21 nt are the predominant species in a 35S Pol II line, whereas 24- and/or 22-nucleotide (nt) siRNAs are produced by a Pol III line. The 35S line accumulated the loop of the hpRNA, in contrast to full-length hpRNA in the Pol III line. These suggest that Pol II and Pol III-transcribed hpRNAs are processed by different pathways. One Pol III transgene produced only 24-nt siRNAs but silenced the target gene efficiently, indicating that the 24-nt siRNAs can direct mRNA degradation; specific cleavage was confirmed by 59 rapid amplification of cDNA ends (RACE). Both Pol II- and Pol III-directed hpRNA transgenes induced cytosine methylation in the target DNA. The extent of methylation is not correlated with the level of 21-nt siRNAs, suggesting that they are not effective inducers of DNA methylation. The promoter of a U6 transgene was significantly methylated, whereas the promoter of the endogenous U6 gene was almost free of cytosine methylation, suggesting that endogenous sequences are more resistant to de novo DNA methylation than are transgene constructs. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 RNA Society.
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|Item Type:||Journal Article|
|Additional Information:||Articles free to read on journal website after 12 months|
|Keywords:||Gene silencing, hpRNA, Promoter, RNA polymerase III, RNA-directed DNA methylation, siRNA, beta glucuronidase, complementary DNA, cytosine, DNA directed RNA polymerase III, messenger RNA, nucleotide, RNA polymerase II, short hairpin RNA, small interfering RNA, plant DNA, plant RNA, Arabidopsis, article, controlled study, DNA methylation, gene amplification, gene function, gene sequence, gene targeting, genetic transcription, nonhuman, phytoene desaturase gene, plant genetics, priority journal, promoter region, protein expression, regulator gene, rice, RNA cleavage, RNA degradation, tomato, transgene, gene, genetics, metabolism, nucleotide sequence, plant, RNA processing, tobacco, transgenic plant, Base Sequence, DNA, Plant, Genes, Plant, Oryza sativa, Plants, Plants, Genetically Modified, Promoter Regions (Genetics), RNA Processing, Post-Transcriptional, RNA, Plant, RNA, Small Interfering, PMCID: PMC2327362|
|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 03:18|
|Last Modified:||29 Jan 2015 05:54|
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