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A novel mutation (c.121‑13T>A) in the polypyrimidine tract of the splice acceptor site of intron 2 causes exon 3 skipping in mitochondrial acetoacetyl-CoA thiolase gene.
Mol Med Rep 2017; 15(6):3879-3884MM

Abstract

Mitochondrial acetoacetyl-CoA thiolase (T2) (gene symbol: ACAT1) deficiency is an autosomal recessive disorder affecting isoleucine catabolism and ketone body utilization. In this study, mutational analysis of an Indian T2-deficient patient revealed a homozygous mutation (c.121‑13T>A) located at the polypyrimidine tract of the splice acceptor site of intron 2, and exon 3 skipping was identified by cDNA analysis using cycloheximide. We made three mutant constructs (c.121‑13T>A, T>C, and T>G substitutions) followed by making a wild-type minigene construct that included an ACAT1 segment from exon 2 to 4 for a splicing experiment. The minigene splicing experiment demonstrated that exon 3 skipping was induced not only by c.121‑13T>A mutation, but also by the other two substitutions. It was difficult to predict the effect of these mutations on splicing using in silico tools, as predictions of different tools were inconsistent with each other. The minigene splicing experiment remains the most reliable method to unravel splicing abnormalities.

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.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 Pediatrics, Kasturba Medical College, Manipal University, Manipal 576104, India.Department of Pediatrics, Kasturba Medical College, Manipal University, Manipal 576104, India.Department of Medical Genetics, Kasturba Medical College, Manipal University, Manipal 576104, India.Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501‑1194, Japan.

Pub Type(s)

Case Reports
Journal Article

Language

eng

PubMed ID

28393214

Citation

Aoyama, Yuka, et al. "A Novel Mutation (c.121‑13T>A) in the Polypyrimidine Tract of the Splice Acceptor Site of Intron 2 Causes Exon 3 Skipping in Mitochondrial acetoacetyl-CoA Thiolase Gene." Molecular Medicine Reports, vol. 15, no. 6, 2017, pp. 3879-3884.
Aoyama Y, Sasai H, Abdelkreem E, et al. A novel mutation (c.121‑13T>A) in the polypyrimidine tract of the splice acceptor site of intron 2 causes exon 3 skipping in mitochondrial acetoacetyl-CoA thiolase gene. Mol Med Rep. 2017;15(6):3879-3884.
Aoyama, Y., Sasai, H., Abdelkreem, E., Otsuka, H., Nakama, M., Kumar, S., ... Fukao, T. (2017). A novel mutation (c.121‑13T>A) in the polypyrimidine tract of the splice acceptor site of intron 2 causes exon 3 skipping in mitochondrial acetoacetyl-CoA thiolase gene. Molecular Medicine Reports, 15(6), pp. 3879-3884. doi:10.3892/mmr.2017.6434.
Aoyama Y, et al. A Novel Mutation (c.121‑13T>A) in the Polypyrimidine Tract of the Splice Acceptor Site of Intron 2 Causes Exon 3 Skipping in Mitochondrial acetoacetyl-CoA Thiolase Gene. Mol Med Rep. 2017;15(6):3879-3884. PubMed PMID: 28393214.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel mutation (c.121‑13T>A) in the polypyrimidine tract of the splice acceptor site of intron 2 causes exon 3 skipping in mitochondrial acetoacetyl-CoA thiolase gene. AU - Aoyama,Yuka, AU - Sasai,Hideo, AU - Abdelkreem,Elsayed, AU - Otsuka,Hiroki, AU - Nakama,Mina, AU - Kumar,Sandeep, AU - Aroor,Shrikiran, AU - Shukla,Anju, AU - Fukao,Toshiyuki, Y1 - 2017/04/04/ PY - 2016/08/02/received PY - 2017/03/10/accepted PY - 2017/4/11/pubmed PY - 2018/3/6/medline PY - 2017/4/11/entrez SP - 3879 EP - 3884 JF - Molecular medicine reports JO - Mol Med Rep VL - 15 IS - 6 N2 - Mitochondrial acetoacetyl-CoA thiolase (T2) (gene symbol: ACAT1) deficiency is an autosomal recessive disorder affecting isoleucine catabolism and ketone body utilization. In this study, mutational analysis of an Indian T2-deficient patient revealed a homozygous mutation (c.121‑13T>A) located at the polypyrimidine tract of the splice acceptor site of intron 2, and exon 3 skipping was identified by cDNA analysis using cycloheximide. We made three mutant constructs (c.121‑13T>A, T>C, and T>G substitutions) followed by making a wild-type minigene construct that included an ACAT1 segment from exon 2 to 4 for a splicing experiment. The minigene splicing experiment demonstrated that exon 3 skipping was induced not only by c.121‑13T>A mutation, but also by the other two substitutions. It was difficult to predict the effect of these mutations on splicing using in silico tools, as predictions of different tools were inconsistent with each other. The minigene splicing experiment remains the most reliable method to unravel splicing abnormalities. SN - 1791-3004 UR - https://www.unboundmedicine.com/medline/citation/28393214/A_novel_mutation__c_121‑13T>A__in_the_polypyrimidine_tract_of_the_splice_acceptor_site_of_intron_2_causes_exon_3_skipping_in_mitochondrial_acetoacetyl_CoA_thiolase_gene_ L2 - http://www.spandidos-publications.com/mmr/15/6/3879 DB - PRIME DP - Unbound Medicine ER -