title: The deafness-associated mitochondrial DNA mutation at position 7445, which affects tRNASer(UCN) precursor processing, has long-range effects on NADH dehydrogenase subunit ND6 gene expression.
abstract: The pathogenetic mechanism of the deafness-associated mitochondrial DNA (mtDNA) T7445C mutation has been investigated in several lymphoblastoid cell lines from members of a New Zealand pedigree exhibiting the mutation in homoplasmic form and from control individuals. We show here that the mutation flanks the 3' end of the tRNASer(UCN) gene sequence and affects the rate but not the sites of processing of the tRNA precursor. This causes an average reduction of approximately 70% in the tRNASer(UCN) level and a decrease of approximately 45% in protein synthesis rate in the cell lines analyzed. The data show a sharp threshold in the capacity of tRNASer(UCN) to support the wild-type protein synthesis rate, which corresponds to approximately 40% of the control level of this tRNA. Strikingly, a 7445 mutation-associated marked reduction has been observed in the level of the mRNA for the NADH dehydrogenase (complex I) ND6 subunit gene, which is located approximately 7 kbp upstream and is cotranscribed with the tRNASer(UCN) gene, with strong evidence pointing to a mechanistic link with the tRNA precursor processing defect. Such reduction significantly affects the rate of synthesis of the ND6 subunit and plays a determinant role in the deafness-associated respiratory phenotype of the mutant cell lines. In particular, it accounts for their specific, very significant decrease in glutamate- or malate-dependent O2 consumption. Furthermore, several homoplasmic mtDNA mutations affecting subunits of NADH dehydrogenase may play a synergistic role in the establishment of the respiratory phenotype of the mutant cells.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, California 91125, USA.
authors.forename: M X
authors.initials: MX
authors.lastname: Guan
authors.forename: J A
authors.initials: JA
authors.lastname: Enriquez
authors.forename: N
authors.initials: N
authors.lastname: Fischel-Ghodsian
authors.forename: R S
authors.initials: RS
authors.lastname: Puranam
authors.forename: C P
authors.initials: CP
authors.lastname: Lin
authors.forename: M A
authors.initials: MA
authors.lastname: Maw
authors.forename: G
authors.initials: G
authors.lastname: Attardi
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002455:Cell Division; D019069:Cell Respiration; D003470:Culture Media; D004272:DNA, Mitochondrial; D003638:Deafness; D005690:Galactose; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009245:NADH Dehydrogenase / Q000235:genetics*; D009520:New Zealand; D009690:Nucleic Acid Conformation; D010375:Pedigree; D014176:Protein Biosynthesis; D012322:RNA Precursors; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger; D012362:RNA, Transfer, Ser; D014407:Tumor Cells, Cultured
chemical_list: D003470:Culture Media; D004272:DNA, Mitochondrial; D012322:RNA Precursors; D012333:RNA, Messenger; D012362:RNA, Transfer, Ser; D009245:NADH Dehydrogenase; D005947:Glucose; D005690:Galactose