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Series GSE146873 Query DataSets for GSE146873
Status Public on Apr 29, 2021
Title RNA m6A methylation of 3ʹ splice site prevents binding of U2AF35 to inhibit splicing
Organisms Caenorhabditis elegans; Bombyx mori; Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary OThe N6-methyladenosine (m6A) RNA modification is widely used to alter the fate of mRNAs. Here we demonstrate that the C. elegans writer METT-10 (orthologue of mouse METTL16) deposits an m6A mark on the 3′ splice site (AG) of the SAM synthetase pre-mRNA which inhibits its proper splicing and protein production. The mechanism is triggered by a rich diet, and acts as an m6A-mediated switch to stop SAM production and regulate its homeostasis. Although the mammalian SAM synthetase pre-mRNA is not regulated via this mechanism, we show that splicing inhibition by 3′ splice site m6A is conserved in mammals. The modification functions by physically preventing the essential splicing factor U2AF35 from recognizing the 3′ splice site. We propose that use of splice site m6A is an ancient mechanism for splicing regulation.
Overall design Abundance and distribution of m6A modification was compared between transcriptomes of the mouse and the worm by m6A-IP RNA-Seq on mixed equimolar amounts of corresponding RNAs. The effect of METT-10 on m6A distribution was analyzed by performing m6A-IP RNA-Seq on RNA isolated from wild-type and mutant METT10 worms grown either on NA22 or OP50 plates. Biological triplicates were used throuhout the study.
Web link
Contributor(s) Mendel M, Delaney K, Raman Pandey R, Chen K, Wenda JM, Broberg Vågbø C, Steiner FA, Homolka D, Pillai RS
Citation(s) 33930289
Submission date Mar 12, 2020
Last update date Jul 02, 2021
Contact name Ramesh Pillai
Organization name University of Geneva
Department Department of Molecular Biology
Street address 30, Quai Ernest-Ansermet
City Gneveva
ZIP/Postal code CH-1211
Country Switzerland
Platforms (3)
GPL19757 Illumina NextSeq 500 (Caenorhabditis elegans)
GPL22765 Illumina HiSeq 4000 (Caenorhabditis elegans)
GPL28266 Illumina NextSeq 500 (Bombyx mori; Caenorhabditis elegans; Mus musculus)
Samples (42)
GSM4408141 RR723_mix_input_total RNA
GSM4408142 RR724_mix_input_total RNA
GSM4408143 RR725_mix_input_total RNA
BioProject PRJNA612262
SRA SRP252561

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Supplementary file Size Download File type/resource
GSE146873_DESeq_NA22_plates.csv.gz 654.9 Kb (ftp)(http) CSV
GSE146873_DESeq_OP50_plates.csv.gz 1.4 Mb (ftp)(http) CSV
GSE146873_MSPC_peaks.xlsx.gz 8.2 Mb (ftp)(http) XLSX
GSE146873_RAW.tar 5.7 Gb (http)(custom) TAR (of BW)
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Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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