Identification of Host Factors Interacting with a γ-Shaped RNA Element from a Plant Virus-Associated Satellite RNA
Open Access
Abstract
Previously, we identified a highly conserved, γ-shaped RNA element (γRE) from satellite RNAs of cucumber mosaic virus (CMV), and we determined γRE to be structurally required for satRNA survival and the inhibition of CMV replication. It remains unknown how γRE biologically functions. In this work, pull-down assays were used to screen candidates of host factors from Nicotiana benthamiana plants using biotin-labeled γRE as bait. Nine host factors were found to interact specifically with γRE. Then, all of these host factors were down-regulated individually in N. benthamiana plants via tobacco rattle virus-induced gene silencing and tested with infection by GFP-expressing CMV (CMV-gfp) and the isolate T1 of satRNA (sat-T1). Out of nine candidates, three host factors, namely histone H3, GTPase Ran3, and eukaryotic translation initiation factor 4A, were extremely important for infection by CMV-gfp and sat-T1. Moreover, we found that cytosolic glyceraldehyde-3-phosphate dehydrogenase 2 contributed to the replication of CMV and sat-T1, but also negatively regulated CMV 2b activity. Collectively, our work provides essential clues for uncovering the mechanism by which satRNAs inhibit CMV replication.Keywords
Funding Information
- National Natural Science Foundation of China (31870144)
This publication has 60 references indexed in Scilit:
- Tobacco calmodulin-like protein provides secondary defense by binding to and directing degradation of virus RNA silencing suppressorsProceedings of the National Academy of Sciences, 2012
- RNA binding is more critical to the suppression of silencing function ofCucumber mosaic virus2b protein than nuclear localizationRNA, 2012
- Glyceraldehyde 3-Phosphate Dehydrogenase Negatively Regulates the Replication of Bamboo Mosaic Virus and Its Associated Satellite RNAJournal of Virology, 2011
- A Viral Satellite RNA Induces Yellow Symptoms on Tobacco by Targeting a Gene Involved in Chlorophyll Biosynthesis using the RNA Silencing MachineryPLOS Pathogens, 2011
- Translation elongation factor 1A is a component of the tombusvirus replicase complex and affects the stability of the p33 replication co-factorVirology, 2009
- Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR dataNucleic Acids Research, 2009
- Statistical methods for efficiency adjusted real‐time PCR quantificationBiotechnology Journal, 2008
- Suppression of Antiviral Silencing by Cucumber Mosaic Virus 2b Protein in Arabidopsis Is Associated with Drastically Reduced Accumulation of Three Classes of Viral Small Interfering RNAsThe Plant Cell, 2007
- Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defenseGenes & Development, 2006
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970