Nanopore direct RNA sequencing maps the complexity of arabidopsis mRNA processing and m6A modification

Parker, Matthew T., Knop, Katarzyna, Sherwood, Anna V., Schurch, Nicholas J., Mackinnon, Katarzyna, Gould, Peter D., Hall, Anthony J. W., Barton, Geoffrey J. and Simpson, Gordon G. (2020) Nanopore direct RNA sequencing maps the complexity of arabidopsis mRNA processing and m6A modification. eLife, 9. ISSN 2050-084X

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Abstract

Understanding genome organization and gene regulation requires insight into RNA transcription, processing and modification. We adapted nanopore direct RNA sequencing to examine RNA from a wild-type accession of the model plant Arabidopsis thaliana and a mutant defective in mRNA methylation (m6A). Here we show that m6A can be mapped in full-length mRNAs transcriptome-wide and reveal the combinatorial diversity of cap-associated transcription start sites, splicing events, poly(A) site choice and poly(A) tail length. Loss of m6A from 3’ untranslated regions is associated with decreased relative transcript abundance and defective RNA 30 end formation. A functional consequence of disrupted m6A is a lengthening of the circadian period. We conclude that nanopore direct RNA sequencing can reveal the complexity of mRNA processing and modification in full-length single molecule reads. These findings can refine Arabidopsis genome annotation. Further, applying this approach to less well-studied species could transform our understanding of what their genomes encode.

Item Type: Article
Uncontrolled Keywords: neuroscience(all),immunology and microbiology(all),biochemistry, genetics and molecular biology(all) ,/dk/atira/pure/subjectarea/asjc/2800
Faculty \ School: Faculty of Science > School of Biological Sciences
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 19 Feb 2021 01:04
Last Modified: 09 Jul 2021 07:17
URI: https://ueaeprints.uea.ac.uk/id/eprint/79284
DOI: 10.7554/eLife.49658

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