Modification Defect at Anticodon Wobble Nucleotide of Mitochondrial tRNAsLeu(UUR) with Pathogenic Mutations of Mitochondrial Myopathy, Encephalopathy, Lactic Acidosis, and Stroke-like Episodes*

The mitochondrial tRNALeu(UUR) (R = A or G) gene possesses several hot spots for pathogenic mutations. A point mutation at nucleotide position 3243 or 3271 is associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes and maternally inherited diabetes with deafness. Detailed studies on two tRNAsLeu(UUR) with the 3243 or 3271 mutation revealed some common characteristics in cybrid cells: (i) a decreased life span, resulting in a 70% decrease in the amounts of the tRNAs in the steady state, (ii) a slight decrease in the ratios of aminoacyl-tRNAsLeu(UUR) versusuncharged tRNAsLeu(UUR), and (iii) accurate aminoacylation with leucine without any misacylation. As a marked result, both of the mutant tRNA molecules were deficient in a modification of uridine that occurs in the normal tRNALeu(UUR) at the first position of the anticodon. The lack of this modification may lead to the mistranslation of leucine into non-cognate phenylalanine codons by mutant tRNAsLeu(UUR), according to the mitochondrial wobble rule, and/or a decrease in the rate of mitochondrial protein synthesis. This finding could explain why two different mutations (3243 and 3271) manifest indistinguishable clinical features.

[1]  N. Bresolin,et al.  In vitro genetic transfer of protein synthesis and respiration defects to mitochondrial DNA-less cells with myopathy-patient mitochondria. , 1991, Molecular and cellular biology.

[2]  U. RajBhandary,et al.  Novel features in the genetic code and codon reading patterns in Neurospora crassa mitochondria based on sequences of six mitochondrial tRNAs. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Y. Kagawa,et al.  Effects of ethidium bromide treatment of mouse cells on expression and assembly of nuclear-coded subunits of complexes involved in the oxidative phosphorylation. , 1990, Biochemical and biophysical research communications.

[4]  K. Watanabe,et al.  7-Methylguanosine at the anticodon wobble position of squid mitochondrial tRNA(Ser)GCU: molecular basis for assignment of AGA/AGG codons as serine in invertebrate mitochondria. , 1998, Biochimica et biophysica acta.

[5]  M. Parisi,et al.  Impairment of mitochondrial transcription termination by a point mutation associated with the MELAS subgroup of mitochondrial encephalomyopathies , 1991, Nature.

[6]  S. Dimauro,et al.  A new mutation associated with MELAS is located in a mitochondrial DNA polypeptide-coding gene , 1995, Neuromuscular Disorders.

[7]  S. Osawa,et al.  Codon recognition patterns as deduced from sequences of the complete set of transfer RNA species in Mycoplasma capricolum. Resemblance to mitochondria. , 1989, Journal of molecular biology.

[8]  T. Ohama,et al.  Translation of synonymous codons in family boxes by Mycoplasma capricolum tRNAs with unmodified uridine or adenosine at the first anticodon position. , 1995, Journal of molecular biology.

[9]  I. Nonaka,et al.  A mutation in the tRNALeu(UUR) gene associated with the MELAS subgroup of mitochondrial encephalomyopathies , 1990, Nature.

[10]  R. Sakuta,et al.  A subtype of diabetes mellitus associated with a mutation of mitochondrial DNA. , 1994, The New England journal of medicine.

[11]  H. Donis-Keller,et al.  Phy M: an RNase activity specific for U and A residues useful in RNA sequence analysis. , 1980, Nucleic acids research.

[12]  K. Nihei,et al.  A point mutation in the mitochondrial tRNA(Leu)(UUR) gene in MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes). , 1990, Biochemical and biophysical research communications.

[13]  N. Hanyu,et al.  Analysis of modified nucleosides and nucleotide sequence of tRNA. , 1987, Methods in enzymology.

[14]  H. Reichmann,et al.  Pathophysiology of the MELAS 3243 Transition Mutation* , 1997, The Journal of Biological Chemistry.

[15]  R. Sakuta,et al.  Accumulation of mtDNA with a mutation at position 3271 in tRNA(Leu)(UUR) gene introduced from a MELAS patient to HeLa cells lacking mtDNA results in progressive inhibition of mitochondrial respiratory function. , 1993, Biochemical and biophysical research communications.

[16]  K. Watanabe,et al.  Higher-order structure of bovine mitochondrial tRNA(Phe) lacking the 'conserved' GG and T psi CG sequences as inferred by enzymatic and chemical probing. , 1994, Nucleic acids research.

[17]  T. Samuelsson,et al.  Unconventional reading of the glycine codons. , 1983, The Journal of biological chemistry.

[18]  Y. Goto,et al.  Clinical Picture of a Case of Diabetes With Mitochondrial tRNA Mutation at Position 3271 , 1996, Diabetes Care.

[19]  T Suzuki,et al.  The 'polysemous' codon--a codon with multiple amino acid assignment caused by dual specificity of tRNA identity. , 1997, The EMBO journal.

[20]  G. Keith,et al.  Mobilities of modified ribonucleotides on two-dimensional cellulose thin-layer chromatography. , 1995, Biochimie.

[21]  F. Sanger,et al.  Different pattern of codon recognition by mammalian mitochondrial tRNAs. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Fenn,et al.  Ultrasonically assisted electrospray ionization for LC/MS determination of nucleosides from a transfer RNA digest. , 1994, Analytical chemistry.

[23]  M. Zeviani,et al.  Complex I deficiency is associated with 3243G:C mitochondrial DNA in osteosarcoma cell cybrids. , 1996, Human molecular genetics.

[24]  I. Nonaka,et al.  MELAS mutation in mtDNA binding site for transcription termination factor causes defects in protein synthesis and in respiration but no change in levels of upstream and downstream mature transcripts. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  U. RajBhandary,et al.  Direct analysis of aminoacylation levels of tRNAs in vivo. Application to studying recognition of Escherichia coli initiator tRNA mutants by glutaminyl-tRNA synthetase. , 1991, The Journal of biological chemistry.

[26]  M. King,et al.  Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. , 1989, Science.

[27]  C. Florentz,et al.  Search for differences in post-transcriptional modification patterns of mitochondrial DNA-encoded wild-type and mutant human tRNALys and tRNALeu(UUR). , 1999, Nucleic acids research.

[28]  J. Enríquez,et al.  MtDNA mutation in MERRF syndrome causes defective aminoacylation of tRNALys and premature translation termination , 1995, Nature Genetics.

[29]  Y. Goto,et al.  A new mtDNA mutation associated with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS). , 1991, Biochimica et biophysica acta.

[30]  K. Watanabe,et al.  A Novel Wobble Rule Found in Starfish Mitochondria , 1998, The Journal of Biological Chemistry.

[31]  S. Dimauro,et al.  Mitochondrial DNA Mutations and Pathogenesis , 1997, Journal of bioenergetics and biomembranes.

[32]  J. Maassen,et al.  Mutation in mitochondrial tRNALeu(UUR) gene in a large pedigree with maternally transmitted type II diabetes mellitus and deafness , 1992, Nature Genetics.

[33]  M. Mather,et al.  Intrinsic uncoupling of cytochrome c oxidase may cause the maternally inherited mitochondrial diseases MELAS and LHON , 1998, FEBS letters.

[34]  S. Dimauro,et al.  A new mtDNA mutation in the tRNA(Lys) gene associated with myoclonic epilepsy and ragged-red fibers (MERRF). , 1992, American journal of human genetics.

[35]  S. Osawa,et al.  Occurrence of unmodified adenine and uracil at the first position of anticodon in threonine tRNAs in Mycoplasma capricolum. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[36]  D. Wallace,et al.  Myoclonic epilepsy and ragged-red fiber disease (MERRF) is associated with a mitochondrial DNA tRNALys mutation , 1990, Cell.

[37]  F. Sanger,et al.  Sequence and organization of the human mitochondrial genome , 1981, Nature.

[38]  K. Nihei,et al.  Respiration-deficient Cells AreCausedbya Single Point Mutation intheMitochondrial tRNA-Leu(UUR)Genein Mitochondrial Myopathy, Encephalopathy, Lactic Acidosis, andStrokelike Episodes (MELAS) , 1991 .

[39]  J. Hayashi,et al.  Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondrial DNA results in mitochondrial dysfunction. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[40]  E. Schon,et al.  Defects in mitochondrial protein synthesis and respiratory chain activity segregate with the tRNA(Leu(UUR)) mutation associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes. , 1992, Molecular and cellular biology.