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Exon 10 skipping in ACAT1 caused by a novel c.949G>A mutation located at an exonic splice enhancer site.
Mol Med Rep 2016; 14(5):4906-4910MM

Abstract

Beta-ketothiolase deficiency, also known as mitochondrial acetoacetyl-CoA thiolase (T2) deficiency, is an autosomal recessive disease caused by mutations in the acetyl‑CoA acetyltransferase 1 (ACAT1) gene. A German T2‑deficient patient that developed a severe ketoacidotic episode at the age of 11 months, was revealed to be a compound heterozygote of a previously reported null mutation, c.472A>G (p.N158D) and a novel mutation, c.949G>A (p.D317N), in ACAT1. The c.949G>A mutation was suspected to cause aberrant splicing as it is located within an exonic splicing enhancer sequence (c. 947CTGACGC) that is a potential binding site for serine/arginine‑rich splicing factor 1. A mutation in this sequence, c.951C>T, results in exon 10 skipping. A minigene construct was synthesized that included exon 9‑truncated intron 9‑exon 10‑truncated intron 10‑exon 11, and the splicing of this minigene revealed that the c.949G>A mutant construct caused exon 10 skipping in a proportion of the transcripts. Furthermore, additional substitution of G for C at the first nucleotide of exon 10 (c.941G>C) abolished the effect of the c.949G>A mutation. Transient expression analysis of the c.949G>A mutant cDNA revealed no residual T2 activity in the mutated D317N enzyme. Therefore, c.949G>A (D317N) is a pathogenic missense mutation, and diminishes the effect of an exonic splicing enhancer and causes exon 10 skipping. The present study demonstrates that a missense mutation, or even a synonymous substitution, may disrupt enzyme function by interference with splicing.

Authors+Show Affiliations

Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.Division of Clinical Genetics, Gifu University Hospital, Gifu 501‑1194, Japan.Department of Biomedical Sciences, College of Life and Health Sciences, Education and Training Center of Medical Technology, Chubu University, Kasugai 487‑8501, Japan.Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.Department of Pediatrics, Medical University of Vienna, Vienna A‑1090, Austria.Bioanalytics and Biochemistry, Department of Natural Sciences, University of Applied Sciences, D‑53359 Rheinbach, Germany.Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.

Pub Type(s)

Case Reports
Journal Article

Language

eng

PubMed ID

27748876

Citation

Otsuka, Hiroki, et al. "Exon 10 Skipping in ACAT1 Caused By a Novel c.949G>A Mutation Located at an Exonic Splice Enhancer Site." Molecular Medicine Reports, vol. 14, no. 5, 2016, pp. 4906-4910.
Otsuka H, Sasai H, Nakama M, et al. Exon 10 skipping in ACAT1 caused by a novel c.949G>A mutation located at an exonic splice enhancer site. Mol Med Rep. 2016;14(5):4906-4910.
Otsuka, H., Sasai, H., Nakama, M., Aoyama, Y., Abdelkreem, E., Ohnishi, H., ... Fukao, T. (2016). Exon 10 skipping in ACAT1 caused by a novel c.949G>A mutation located at an exonic splice enhancer site. Molecular Medicine Reports, 14(5), pp. 4906-4910. doi:10.3892/mmr.2016.5819.
Otsuka H, et al. Exon 10 Skipping in ACAT1 Caused By a Novel c.949G>A Mutation Located at an Exonic Splice Enhancer Site. Mol Med Rep. 2016;14(5):4906-4910. PubMed PMID: 27748876.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Exon 10 skipping in ACAT1 caused by a novel c.949G>A mutation located at an exonic splice enhancer site. AU - Otsuka,Hiroki, AU - Sasai,Hideo, AU - Nakama,Mina, AU - Aoyama,Yuka, AU - Abdelkreem,Elsayed, AU - Ohnishi,Hidenori, AU - Konstantopoulou,Vassiliki, AU - Sass,Jörn Oliver, AU - Fukao,Toshiyuki, Y1 - 2016/10/10/ PY - 2016/06/04/received PY - 2016/08/25/accepted PY - 2016/10/26/pubmed PY - 2017/4/7/medline PY - 2016/10/26/entrez SP - 4906 EP - 4910 JF - Molecular medicine reports JO - Mol Med Rep VL - 14 IS - 5 N2 - Beta-ketothiolase deficiency, also known as mitochondrial acetoacetyl-CoA thiolase (T2) deficiency, is an autosomal recessive disease caused by mutations in the acetyl‑CoA acetyltransferase 1 (ACAT1) gene. A German T2‑deficient patient that developed a severe ketoacidotic episode at the age of 11 months, was revealed to be a compound heterozygote of a previously reported null mutation, c.472A>G (p.N158D) and a novel mutation, c.949G>A (p.D317N), in ACAT1. The c.949G>A mutation was suspected to cause aberrant splicing as it is located within an exonic splicing enhancer sequence (c. 947CTGACGC) that is a potential binding site for serine/arginine‑rich splicing factor 1. A mutation in this sequence, c.951C>T, results in exon 10 skipping. A minigene construct was synthesized that included exon 9‑truncated intron 9‑exon 10‑truncated intron 10‑exon 11, and the splicing of this minigene revealed that the c.949G>A mutant construct caused exon 10 skipping in a proportion of the transcripts. Furthermore, additional substitution of G for C at the first nucleotide of exon 10 (c.941G>C) abolished the effect of the c.949G>A mutation. Transient expression analysis of the c.949G>A mutant cDNA revealed no residual T2 activity in the mutated D317N enzyme. Therefore, c.949G>A (D317N) is a pathogenic missense mutation, and diminishes the effect of an exonic splicing enhancer and causes exon 10 skipping. The present study demonstrates that a missense mutation, or even a synonymous substitution, may disrupt enzyme function by interference with splicing. SN - 1791-3004 UR - https://www.unboundmedicine.com/medline/citation/27748876/Exon_10_skipping_in_ACAT1_caused_by_a_novel_c_949G>A_mutation_located_at_an_exonic_splice_enhancer_site_ L2 - http://www.spandidos-publications.com/mmr/14/5/4906 DB - PRIME DP - Unbound Medicine ER -