Contract grant sponsors: The Danish Medical Research Council (271-07-342, 11-107174, 271-08-0120); The Novo Nordisk Foundation (15430).
The ETFDH c.158A>G Variation Disrupts the Balanced Interplay of ESE- and ESS-Binding Proteins thereby Causing Missplicing and Multiple Acyl-CoA Dehydrogenation Deficiency
Article first published online: 28 OCT 2013
© 2013 WILEY PERIODICALS, INC.
Volume 35, Issue 1, pages 86–95, January 2014
How to Cite
Olsen, R. K. J., Brøner, S., Sabaratnam, R., Doktor, T. K., Andersen, H. S., Bruun, G. H., Gahrn, B., Stenbroen, V., Olpin, S. E., Dobbie, A., Gregersen, N. and Andresen, B. S. (2014), The ETFDH c.158A>G Variation Disrupts the Balanced Interplay of ESE- and ESS-Binding Proteins thereby Causing Missplicing and Multiple Acyl-CoA Dehydrogenation Deficiency. Hum. Mutat., 35: 86–95. doi: 10.1002/humu.22455
Communicated by Peter K. Rogan
- Issue published online: 18 DEC 2013
- Article first published online: 28 OCT 2013
- Accepted manuscript online: 7 OCT 2013 10:47AM EST
- Manuscript Accepted: 25 SEP 2013
- Manuscript Received: 19 DEC 2012
- The Danish Medical Research Council. Grant Numbers: 271-07-342, 11-107174, 271-08-0120
- The Novo Nordisk Foundation. Grant Number: 15430
- exonic splicing enhancer;
- exonic splicing silencer;
- pre-mRNA splicing;
Multiple acyl-CoA dehydrogenation deficiency is a disorder of fatty acid and amino acid oxidation caused by defects of electron transfer flavoprotein (ETF) or its dehydrogenase (ETFDH). A clear relationship between genotype and phenotype makes genotyping of patients important not only diagnostically but also for prognosis and for assessment of treatment. In the present study, we show that a predicted benign ETFDH missense variation (c.158A>G/p.Lys53Arg) in exon 2 causes exon skipping and degradation of ETFDH protein in patient samples. Using splicing reporter minigenes and RNA pull-down of nuclear proteins, we show that the c.158A>G variation increases the strength of a preexisting exonic splicing silencer (ESS) motif UAGGGA. This ESS motif binds splice inhibitory hnRNP A1, hnRNP A2/B1, and hnRNP H proteins. Binding of these inhibitory proteins prevents binding of the positive splicing regulatory SRSF1 and SRSF5 proteins to nearby and overlapping exonic splicing enhancer elements and this causes exon skipping. We further suggest that binding of hnRNP proteins to UAGGGA is increased by triggering synergistic hnRNP H binding to GGG triplets located upstream and downsteam of the UAGGGA motif. A number of disease-causing exonic elements that induce exon skipping in other genes have a similar architecture as the one in ETFDH exon 2.