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PMID: 28977517 Published · ppublish English Journal Article

N6-methyladenosine demethylase FTO targets pre-mRNAs and regulates alternative splicing and 3'-end processing.

Nucleic acids research ·Vol. 45 ·No. 19 ·2017-11-02 ·Pages 11356-11370

Bartosovic M, Molares HC, Gregorova P, Hrossova D, Kudla G, Vanacova S

Abstract

N6-methyladenosine (m6A) is the most abundant base modification found in messenger RNAs (mRNAs). The discovery of FTO as the first m6A mRNA demethylase established the concept of reversible RNA modification. Here, we present a comprehensive transcriptome-wide analysis of RNA demethylation and uncover FTO as a potent regulator of nuclear mRNA processing events such as alternative splicing and 3΄ end mRNA processing. We show that FTO binds preferentially to pre-mRNAs in intronic regions, in the proximity of alternatively spliced (AS) exons and poly(A) sites. FTO knockout (KO) results in substantial changes in pre-mRNA splicing with prevalence of exon skipping events. The alternative splicing effects of FTO KO anti-correlate with METTL3 knockdown suggesting the involvement of m6A. Besides, deletion of intronic region that contains m6A-linked DRACH motifs partially rescues the FTO KO phenotype in a reporter system. All together, we demonstrate that the splicing effects of FTO are dependent on the catalytic activity in vivo and are mediated by m6A. Our results reveal for the first time the dynamic connection between FTO RNA binding and demethylation activity that influences several mRNA processing events.

MeSH Terms
3' Untranslated Regions/genetics Adenosine/analogs & derivatives,metabolism Alpha-Ketoglutarate-Dependent Dioxygenase FTO/genetics,metabolism Alternative Splicing Exons/genetics Gene Expression Profiling/methods HEK293 Cells Humans Introns/genetics Methyltransferases/genetics,metabolism Mutagenesis, Site-Directed Mutation Poly A/genetics Protein Binding RNA Precursors/genetics,metabolism
Chemicals
3' Untranslated Regions RNA Precursors Poly A N-methyladenosine Alpha-Ketoglutarate-Dependent Dioxygenase FTO Methyltransferases METTL3 protein, human Adenosine
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Bartosovic Marek
CEITEC-Central European Institute of Technology, Masaryk University, Brno 62500, Czech Republic.
Molares Helena Covelo
CEITEC-Central European Institute of Technology, Masaryk University, Brno 62500, Czech Republic.
Gregorova Pavlina
CEITEC-Central European Institute of Technology, Masaryk University, Brno 62500, Czech Republic.
Hrossova Dominika
CEITEC-Central European Institute of Technology, Masaryk University, Brno 62500, Czech Republic.
Kudla Grzegorz
MRC Human Genetics Unit MRC IGMM, University of Edinburgh Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK.
Vanacova Stepanka
CEITEC-Central European Institute of Technology, Masaryk University, Brno 62500, Czech Republic.
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkx778
Published
2017-11-02
Pages
11356-11370
Language
English
Region
England
NLM ID
0411011
PMCID
PMC5737695
Subset
IM
Grants
Medical Research Council · MC_UU_12018/23 · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/gkx778 PMC Full Text View journal details
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