Abstract
A block of single-nucleotide polymorphisms within intron 1 of the FTO (fat mass and obesity associated) gene is associated with variation in body weight. Previous works suggest that increased expression of FTO, which encodes a 2-oxoglutarate-dependent nucleic acid demethylase, leads to increased body weight, although the underlying mechanism has remained unclear. To elucidate the function of FTO, we examined the consequences of altered FTO levels in cultured cells and murine brain. Here we show that a knockdown of FTO in HEK293 cells affects the transcripts levels of genes involved in the response to starvation, whereas overexpression of FTO affects the transcript levels of genes related to RNA processing and metabolism. Subcellular localization of FTO further strengthens the latter notion. Using immunocytochemistry and confocal laser scanning microscopy, we detected FTO in nuclear speckles and – to a lesser and varying extent – in the nucleoplasm and nucleoli of HEK293, HeLa and MCF-7 cells. Moreover, RNA modification analyses revealed that loss of Fto affects the 3-methyluridine/uridine and pseudouridine/uridine ratios in total brain RNA. We conclude that altered levels of FTO have multiple and diverse consequences on RNA modifications and the transcriptome.
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Acknowledgements
We thank Professor G Iliakis for providing access to the confocal laser microscope and Professor M Linscheid for providing equipment and helpful suggestions. We are grateful to one of the anonymous reviewers for pointing out the genes related to ciliary function. We are thankful to Christian Grosser for sharing his data. This work was supported by the Bundesministerium für Bildung und Forschung (NGFN plus 01GS0820) and Deutsche Forschungsgemeinschaft (LI309/19-1, LI309/30-2).
Accession code: Miccroarray data generated by hybridization of RNA from both FTO1_C1 and FTO2_D4 clones with and without induced expression as well as from FTO siRNA- and scrampled siRNA-treansfected cells can be obtained from Gene Expression Omnibus (GSE33870). URL: GeneTrail gene set analysis tool at http://genetrail.bioinf.uni-sb.de/).
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Berulava, T., Ziehe, M., Klein-Hitpass, L. et al. FTO levels affect RNA modification and the transcriptome. Eur J Hum Genet 21, 317–323 (2013). https://doi.org/10.1038/ejhg.2012.168
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DOI: https://doi.org/10.1038/ejhg.2012.168
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