Journal article
The presence of high-molecular-weight viral RNAs interferes with the detection of viral small RNAs
RNA, Vol.16(5), pp.1062-1067
2010
PMCID: PMC2856878
PMID: 20348444
Abstract
Viral small interfering RNA (siRNA) accumulation in plants is reported to exhibit a strong strand polarity bias, with plus (+) strand siRNAs dominating over minus (−) strand populations. This is of particular interest, as siRNAs processed from double-stranded RNA would be expected to accumulate equivalent amounts of both species. Here, we show that, as reported, (−) strand viral siRNAs are detected at much lower levels than (+) strand-derived species using standard Northern hybridization approaches. However, when total RNA is spiked with in vitro-transcribed antisense viral genomic RNA, (−) strand viral siRNAs are detected at increased levels equivalent to those of (+) strand siRNA. Our results suggest that (+) and (−) strand viral siRNAs accumulate to equivalent levels; however, a proportion of the (−) strand siRNAs are sequestered from the total detectable small RNA population during gel electrophoresis by hybridizing to the high-molecular-weight sense strand viral genomic RNA. Our findings provide a plausible explanation for the observed strand bias of viral siRNA accumulation, and could have wider implications in the analysis of both viral and nonviral small RNA accumulation.
Details
- Title
- The presence of high-molecular-weight viral RNAs interferes with the detection of viral small RNAs
- Authors
- Neil A. Smith (Author) - CSIRO Plant IndustryAndrew Eamens (Author) - CSIRO Plant IndustryMing-Bo Wang (Author) - CSIRO Plant Industry
- Publication details
- RNA, Vol.16(5), pp.1062-1067
- Publisher
- Cold Spring Harbor Laboratory Press
- DOI
- 10.1261/rna.2049510
- ISSN
- 1469-9001
- PMID
- 20348444; PMC2856878
- Organisation Unit
- University of the Sunshine Coast, Queensland; School of Health and Behavioural Sciences - Legacy; School of Health - Biomedicine
- Language
- English
- Record Identifier
- 99656589702621
- Output Type
- Journal article
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