title: Functional and morphological abnormalities of mitochondria harbouring the tRNA(Leu)(UUR) mutation in mitochondrial DNA derived from patients with maternally inherited diabetes and deafness (MIDD) and progressive kidney disease.
abstract: OBJECTIVE
  An A to G transition at nucleotide position 3243 in the mitochondrial tRNA Leu(UUR) gene has been identified in patients with maternally inherited diabetes and deafness, as well as in patients with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes, chronic progressive external ophpthalmoplegia, cardiomyopathy and progressive kidney disease. Variations in the mitochondrial DNA haplotype as well as differences in the degree and distribution of heteroplasmy in a certain tissue are factors that may contribute to the variety in phenotypical expression of the 3243 tRNA(Leu)(UUR) mutation. We have done morphological and functional experiments on mitochondria carrying the 3243 mutation derived from patients with either maternally inherited diabetes and deafness or progressive kidney disease to prove the pathogenicity of the 3243 mutation and to examine whether the mtDNA haplotype modulates the pathobiochemistry of this mutation.
  
  
  METHODS
  We constructed clonal cell lines that contain predominantly mutated or exclusively wild-type mtDNA with a distinct mtDNA haplotype by the methodology of mitochondria-mediated transformation. Cells lacking mitochondrial DNA (rho(o)) were used as recipients and donor mitochondria were derived from fibroblasts of a patient with either maternally inherited diabetes and deafness or progressive kidney disease. The fibroblasts from these clinically distinct patients carry different mitochondrial DNA haplotypes with the 3243 mutation in heteroplasmic form.
  
  
  RESULTS
  Heteroplasmy in the clonal cybrid cells ranged from 0 to 100%, reflecting the heterogeneity of the mitochondrial donor cell. Cybrid cells containing predominantly mutant mitochondrial DNA showed lactic acidosis, poor respiration and marked defects in mitochondrial morphology and respiratory chain complex I and IV activities. No differences were observed in the extent of the mitochondrial dysfunction between the mutant cells derived from the two donors.
  
  
  CONCLUSIONS
  These results provide evidence for a pathogenic effect of the tRNA(Leu)(UUR) mutation in maternally inherited diabetes and deafness and progressive kidney disease, and show no evidence of a contribution of the mitochondrial DNA haplotype as a modulating the biochemical expression of the mutation.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Centre, The Netherlands.
authors.forename: J M
authors.initials: JM
authors.lastname: van den Ouweland
authors.forename: P
authors.initials: P
authors.lastname: Maechler
authors.forename: C B
authors.initials: CB
authors.lastname: Wollheim
authors.forename: G
authors.initials: G
authors.lastname: Attardi
authors.forename: J A
authors.initials: JA
authors.lastname: Maassen
mesh_terms: D002460:Cell Line; D004272:DNA, Mitochondrial / Q000235:genetics*; D003638:Deafness / Q000235:genetics*; D003924:Diabetes Mellitus, Type 2 / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D006239:Haplotypes; D006801:Humans; D007674:Kidney Diseases / Q000235:genetics*; D009154:Mutation; D010101:Oxygen Consumption; D010375:Pedigree; D010641:Phenotype; D012356:RNA, Transfer, Leu / Q000235:genetics*; D013577:Syndrome
chemical_list: D004272:DNA, Mitochondrial; D012356:RNA, Transfer, Leu