Heterologous expression of plant virus genes that suppress post-transcriptional gene silencing results in suppression of RNA interference in Drosophila cells

BMC Biotechnology
Brian ReavyStuart A MacFarlane

Abstract

RNA interference (RNAi) in animals and post-transcriptional gene silencing (PTGS) in plants are related phenomena whose functions include the developmental regulation of gene expression and protection from transposable elements and viruses. Plant viruses respond by expressing suppressor proteins that interfere with the PTGS system. Here we demonstrate that both transient and constitutive expression of the Tobacco etch virus HC-Pro silencing suppressor protein, which inhibits the maintenance of PTGS in plants, prevents dsRNA-induced RNAi of a lacZ gene in cultured Drosophila cells. Northern blot analysis of the RNA present in Drosophila cells showed that HC-Pro prevented degradation of lacZ RNA during RNAi but that there was accumulation of the short (23nt) RNA species associated with RNAi. A mutant HC-Pro that does not suppress PTGS in plants also does not affect RNAi in Drosophila. Similarly, the Cucumber mosaic virus 2b protein, which inhibits the systemic spread of PTGS in plants, does not suppress RNAi in Drosophila cells. In addition, we have used the Drosophila system to demonstrate that the 16K cysteine-rich protein of Tobacco rattle virus, which previously had no known function, is a silencing suppressor protein. These ...Continue Reading

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Citations

Sep 3, 2013·Indian Journal of Virology : an Official Organ of Indian Virological Society·Kathy La Fauce, Leigh Owens
May 30, 2006·Journal of Virology·Zsuzsanna MéraiDániel Silhavy
Nov 16, 2010·Biochemistry. Biokhimii︠a︡·R T Omarov, R I Bersimbai
Jun 14, 2006·Annual Review of Microbiology·Feng Li, Shou-Wei Ding
Aug 23, 2006·Nature Immunology·Sara Cherry, Neal Silverman
Jul 14, 2010·Molecular Plant Pathology·Stuart A Macfarlane
May 1, 2007·Plant Disease·Leslie L DomierGlen L Hartman

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Methods Mentioned

BETA
transfection
transfect
transgenic
PCR
electrophoresis

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