Alternative splicing in Acad8 resulting a mitochondrial defect and progressive hepatic steatosis in mice.
Pediatr Res
; 70(1): 31-6, 2011 Jul.
Article
em En
| MEDLINE
| ID: mdl-21659959
ABSTRACT
Using a combination of N-ethyl-N-nitrosourea-mediated mutagenesis and metabolomics-guided screening, we identified mice with elevated blood levels of short-chain C4-acylcarnitine and increased urine isobutyryl-glycine. Genome-wide homozygosity screening, followed by fine mapping, located the disease gene to 15-25 Mb of mouse chromosome 9 where a candidate gene, Acad8, encoding mitochondrial isobutyryl-CoA dehydrogenase was located. Genomic DNA sequencing revealed a single-nucleotide mutation at -17 of the first intron of Acad8 in affected mice. cDNA sequencing revealed an intronic 28-bp insertion at the site of the mutation, which caused a frame shift with a premature stop codon. In vitro splicing assay confirmed that the mutation was sufficient to activate an upstream, aberrant 3' splice site. There was a reduction in the expression of Acad8 at both the mRNA and protein levels. The mutant mice grew normally but demonstrated cold intolerance at young age with a progressive hepatic steatosis. Homozygous mutant mice hepatocytes had abnormal mitochondria with crystalline inclusions, suggestive of mitochondriopathy. This mouse model of isobutyryl-CoA dehydrogenase deficiency could provide us a better understanding of the possible role of IBD deficiency in mitochondriopathy and fatty liver.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Mitocôndrias Hepáticas
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Processamento Alternativo
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Oxirredutases atuantes sobre Doadores de Grupo CH-CH
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Fígado Gorduroso
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Erros Inatos do Metabolismo dos Aminoácidos
Idioma:
En
Revista:
Pediatr Res
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Taiwan