E-GEOD-41432 - Transcription profiling by high throughput sequencing of Arabidopsis upf1, upf3 double mutant and cycloheximide-treated seedlings

Released on 1 October 2013, last updated on 15 April 2016
Arabidopsis thaliana
Samples (12)
Protocols (3)
Nonsense-mediated mRNA decay (NMD) is a eukaryotic surveillance pathway that recognizes aberrant transcripts arising from mutations and transcriptional mistakes. Moreover, differential transcript processing such as alternative precursor mRNA splicing (AS) can generate NMD substrates, however, the extent of coupled AS and NMD remained unclear. To investigate NMD targeting of AS variants, we performed transcriptome-wide splicing studies using Arabidopsis thaliana single and double mutants in the NMD factor homologues UPF1 and UPF3, as well as samples treated with the translation inhibitor cycloheximide (CHX). Our analyses revealed that at least 17.5% of all multi-exon, protein-coding genes produce splicing variants of all major types that are targeted by NMD, with a significant fraction originating from the splicing of cryptic introns. Furthermore, accumulation of many intron-retained mRNAs in the mutants, but not in response to CHX suggests the action of distinct routes of NMD with variable impact of translation. Importantly, 92.3% of the NMD-responsive mRNAs exhibit classical NMD-eliciting features, supporting their authenticity as direct targets. NMD-dependent AS variants are linked to diverse biological functions, including the poison exon-mediated regulation of signaling and posttranslational protein modification components. In addition to mRNAs, numerous non-coding RNAs as well as newly identified transcripts derived from intergenic regions were shown to be NMD responsive. In summary, our comprehensive analysis of AS-coupled NMD provides strong evidence for a major function of this pathway in shaping the transcriptome, having fundamental implications in gene regulation and quality control of transcript processing steps in higher eukaryotes. Comparison of gene expression and alternative splicing patterns in control and nonsense-mediated decay (NMD)-impaired Arabidopsis seedlings
Experiment type
RNA-seq of coding RNA 
Andre Kahles, Andreas Wachter, Anil K Kesarwani, Eva Stauffer, Gabriele Drechsel, Gunnar Rätsch, Jonas Behr, Philipp Drewe
Exp. designProtocolsVariablesProcessedSeq. reads
Investigation descriptionE-GEOD-41432.idf.txt
Sample and data relationshipE-GEOD-41432.sdrf.txt
Additional data (1)E-GEOD-41432.additional.1.zip