Comprehensive cDNA study and quantitative analysis of mutant HADHA and HADHB transcripts in a French cohort of 52 patients with mitochondrial trifunctional protein deficiency

Mol Genet Metab. 2011 Aug;103(4):341-8. doi: 10.1016/j.ymgme.2011.04.006. Epub 2011 Apr 19.

Abstract

Background: Deficiency of mitochondrial trifunctional protein (MTP) is caused by mutations in the HADHA and HADHB genes, which have been mostly delineated at the genomic DNA level and have not been always elucidated.

Aim: To identify mutations in a French cohort of 52 MTP deficient patients and the susceptibility of mutations generating premature termination codons (PTCs) to the nonsense mRNA mediated decay (NMD).

Methods: Mutation screening in fibroblasts was performed at the cDNA level and real-time RT-PCR was used to compare the levels of the different PTC-bearing mRNAs before and after a treatment of fibroblasts by emetine, a translation inhibitor.

Results: A mutation detection rate of 100% was achieved. A total of 22 novel mutations were identified, including a large-sized genomic deletion in HADHB gene. A high proportion of all identified mutations were non-sense, frameshift and splicing mutations, generating (PTCs), distributed essentially on HADHA coding regions. We could demonstrate that the majority of mutations resulting in PTCs conform to the established rules governing the susceptibility to NMD.

Conclusion: Our results emphasize the value of cDNA analysis in the characterization of HADHA and HADHB mutations and further strengthen the model of haploinsufficiency as a major pathomechanism in MTP defects.

MeSH terms

  • Base Sequence
  • Cohort Studies
  • DNA, Complementary / genetics*
  • Female
  • France
  • Haploinsufficiency
  • Humans
  • Lipid Metabolism Disorders / genetics*
  • Male
  • Mitochondrial Diseases / genetics*
  • Mitochondrial Proteins / genetics*
  • Mitochondrial Trifunctional Protein
  • Mitochondrial Trifunctional Protein, alpha Subunit
  • Mitochondrial Trifunctional Protein, beta Subunit
  • Molecular Sequence Data
  • Multienzyme Complexes / genetics*
  • Mutation*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary
  • Mitochondrial Proteins
  • Multienzyme Complexes
  • HADHA protein, human
  • Mitochondrial Trifunctional Protein, alpha Subunit
  • HADHB protein, human
  • Mitochondrial Trifunctional Protein
  • Mitochondrial Trifunctional Protein, beta Subunit