A new mechanism for mtDNA pathogenesis: impairment of post-transcriptional maturation leads to severe depletion of mitochondrial tRNASer(UCN) caused by T7512C and G7497A point mutations

Mollers, M.

doi:10.1093/nar/gki876

Cited by 31Mitochondrial Function and PathologyRNA and protein synthesis mechanismsRNA modifications and cancer

A new mechanism for mtDNA pathogenesis: impairment of post-transcriptional maturation leads to severe depletion of mitochondrial tRNASer(UCN) caused by T7512C and G7497A point mutations

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Mollers, M.·2005·Nucleic Acids Research

DOI 10.1093/nar/gki876PMC PMC1240115

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Abstract

We have studied the consequences of two homoplasmic, pathogenic point mutations (T7512C and G7497A) in the tRNA(Ser(UCN)) gene of mitochondrial (mt) DNA using osteosarcoma cybrids. We identified a severe reduction of tRNA(Ser(UCN)) to levels below 10% of controls for both mutations, resulting in a 40% reduction in mitochondrial protein synthesis rate and in a respiratory chain deficiency resembling that in the patients muscle. Aminoacylation was apparently unaffected. On non-denaturating northern blots we detected an altered electrophoretic mobility for G7497A containing tRNA molecules suggesting a structural impact of this mutation, which was confirmed by structural probing. By comparing in vitro transcribed molecules with native RNA in such gels, we also identified tRNA(Ser(UCN)) being present in two isoforms in vivo, probably corresponding to the nascent, unmodified transcripts co-migrating with the in vitro transcripts and a second, faster moving isoform corresponding to the mature tRNA. In cybrids containing either mutations the unmodified isoforms were severely reduced. We hypothesize that both mutations lead to an impairment of post-transcriptional modification processes, ultimately leading to a preponderance of degradation by nucleases over maturation by modifying enzymes, resulting in severely reduced tRNA(Ser(UCN)) steady state levels. We infer that an increased degradation rate, caused by disturbance of tRNA maturation and, in the case of the G7497A mutant, alteration of tRNA structure, is a new pathogenic mechanism of mt tRNA point mutations.

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m.7512T N C causes slight localised kinking of the acceptor stem.
“The mt-tRNA Ser(UCN) m.7512T N C mutation causes slight localised kinking of the acceptor stem [199]”
From Mitochondrial DNA mutations and human disease (2010) · cites this paper as [199]

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Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961 · see citing works
Molecular Cloning: A Laboratory Manual
Sambrook, J., Fritsch, E., & Maniatis, T. (1989) · UCLA - Biblioteca de Ciencias de la Salud · Cited by 133,589 · see citing works
Molecular cloning: A laboratory manual
Deininger, P. L. (1990) · Analytical Biochemistry · doi:10.1016/0003-2697(90)90595-z · Cited by 84,684 · see citing works
Short-Term Effects of Nose-Only Cigarette Smoke Exposure on Glutathione Redox Homeostasis, Cytochrome P450 1A1/2 and Respiratory Enzyme Activities in Mice Tissues
Raza, H., John, A., & Nemmar, A. (2013) · Cellular Physiology and Biochemistry · doi:10.1159/000350087 · Cited by 47,311 · see citing works
Human Cells Lacking mtDNA: Repopulation with Exogenous Mitochondria by Complementation
King, M. P., & Attardi, G. M. (1989) · Science · doi:10.1126/science.2814477 · Cited by 1,659 · see citing works
Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation
(1990) · Trends in Genetics · doi:10.1016/0168-9525(90)90040-d · Cited by 1,386 · see citing works
Biochemical and molecular investigations in respiratory chain deficiencies
Rustin, P., Chrétien, D., Bourgeron, T., Gérard, B., Rötig, A., Saudubray, J., & Münnich, A. (1994) · Clinica Chimica Acta · doi:10.1016/0009-8981(94)90055-8 · Cited by 1,239 · see citing works
RIBONUCLEASE P: Unity and Diversity in a tRNA Processing Ribozyme
Frank, D. N., & Pace, N. R. (1998) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.67.1.153 · Cited by 465 · see citing works
Search for characteristic structural features of mammalian mitochondrial tRNAs
Helm, M., Brulé, H., FRIEDE, D., et al. (2000) · RNA · doi:10.1017/s1355838200001047 · Cited by 284 · see citing works
Modification Defect at Anticodon Wobble Nucleotide of Mitochondrial tRNAsLeu(UUR) with Pathogenic Mutations of Mitochondrial Myopathy, Encephalopathy, Lactic Acidosis, and Stroke-like Episodes
Yasukawa, T., Suzuki, T., Suzuki, T., Ueda, T., Ohta, S., & Watanabe, K. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.6.4251 · Cited by 268 · see citing works
Human mitochondrial tRNAs in health and disease
Florentz, C., Sohm, B., Tóth, P., P�tz, J., & Sissler, M. (2003) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-003-2343-1 · Cited by 249 · see citing works
Wobble modification defect in tRNA disturbs codon–anticodon interaction in a mitochondrial disease
Yasukawa, T., Suzuki, T., Ishii, N., Ohta, S., & Watanabe, K. (2001) · The EMBO Journal · doi:10.1093/emboj/20.17.4794 · Cited by 240 · see citing works
The presence of modified nucleotides is required for cloverleaf folding of a human mitochondrial tRNA
Helm, M., Brulé, H., Degoul, F., Cepanec, C., Leroux, J. P., Giegé, R., & Florentz, C. (1998) · Nucleic Acids Research · doi:10.1093/nar/26.7.1636 · Cited by 230 · see citing works
Mitochondria-specific RNA-modifying Enzymes Responsible for the Biosynthesis of the Wobble Base in Mitochondrial tRNAs
Umeda, N., Suzuki, T., Yukawa, M., Ohya, Y., Shindo, H., Watanabe, K., & Suzuki, T. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m409306200 · Cited by 230 · see citing works
[18] In vivo labeling and analysis of human mitochondrial translation products
Chomyn, A. (1996) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(96)64020-8 · Cited by 207 · see citing works
The mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episode syndrome-associated human mitochondrial tRNALeu(UUR) mutation causes aminoacylation deficiency and concomitant reduced association of mRNA with ribosomes
Chomyn, A., Enriquez, J. A., Micol, V., Fernandez-Silva, P., & Attardi, G. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m908734199 · Cited by 197 · see citing works
Multiple neonatal deaths due to a homoplasmic mitochondrial DNA mutation
McFarland, R., Clark, K. M., Morris, A. A. M., Taylor, R. W., Macphail, S., Lightowlers, R. N., & Turnbull, D. M. (2002) · Nature Genetics · doi:10.1038/ng819 · Cited by 170 · see citing works
Abnormal RNA processing associated with a novel tRNA mutation in mitochondrial DNA. A potential disease mechanism.
Bindoff, L. A., Howell, N., Poulton, J., McCullough, D. A., Morten, K., Lightowlers, R. N., Turnbull, D. M., & Weber, K. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)36552-4 · Cited by 166 · see citing works
Mitochondrial tRNA 3' end metabolism and human disease
Levinger, L. (2004) · Nucleic Acids Research · doi:10.1093/nar/gkh884 · Cited by 152 · see citing works
Ribozyme processed tRNA transcripts with unfriendly internal promoter for T7 RNA polymerase: production and activity
Fechter, P., Rudinger, J., Giegé, R., et al. (1998) · FEBS Letters · doi:10.1016/s0014-5793(98)01096-5 · Cited by 141 · see citing works
[41 ] Use of polarography to detect respiration defects in cell cultures
Hofhaus, G., Shakeley, R. M., & Attardi, G. M. (1996) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(96)64043-9 · Cited by 139 · see citing works
Defect in modification at the anticodon wobble nucleotide of mitochondrial tRNA<sup>Lys</sup> with the MERRF encephalomyopathy pathogenic mutation
Yasukawa, T., Suzuki, T., Ishii, N., Ueda, T., Ohta, S., & Watanabe, K. (2000) · FEBS Letters · doi:10.1016/s0014-5793(00)01145-5 · Cited by 124 · see citing works
Molecular analysis of cloned human 18S ribosomal DNA segments.
Wilson, G. N., Hollar, B. A. M., Waterson, J. R., & Schmickel, R. D. (1978) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.75.11.5367 · Cited by 119 · see citing works
Progressive myoclonus epilepsy and mitochondrial myopathy associated with mutations in the tRNAser(UCN) gene
Jaksch, M., Klopstock, T., Kurlemann, G., et al. (1998) · Annals of Neurology · doi:10.1002/ana.410440409 · Cited by 110 · see citing works
Complex I deficiency is Associated with 3243G:C Mitochondrial DNA in Osteosarcoma Cell Cybrids
Dunbar, D. R., Moonie, P., Zeviani, M., & Holt, I. J. (1996) · Human Molecular Genetics · doi:10.1093/hmg/5.1.123 · Cited by 107 · see citing works
Impact of disease-related mitochondrial mutations on tRNA structure and function
Wittenhagen, L. M. (2003) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2003.09.006 · Cited by 102 · see citing works
A tRNA suppressor mutation in human mitochondria
Meziane, A. E., Lehtinen, S. K., Hance, N., Nijtmans, L. G. J., Dunbar, D. R., Holt, I. J., & Jacobs, H. T. (1998) · Nature Genetics · doi:10.1038/ng0498-350 · Cited by 101 · see citing works
Towards Understanding Human Mitochondrial Leucine Aminoacylation Identity
Sohm, B., Frugier, M., Brulé, H., Olszak, K., Przykorska, A., & Florentz, C. (2003) · Journal of Molecular Biology · doi:10.1016/s0022-2836(03)00373-5 · Cited by 92 · see citing works
A simple method for 3'-labeling of RNA
Huang, Z. (1996) · Nucleic Acids Research · doi:10.1093/nar/24.21.4360 · Cited by 89 · see citing works
Mitochondrial DNA and RNA processing in MELAS
Kaufmann, P., Shanske, S., Hirano, M., DiMauro, S., King, M. P., Koga, Y., & Schon, E. A. (1996) · Annals of Neurology · doi:10.1002/ana.410400208 · Cited by 75 · see citing works
A novel cloverleaf structure found in mammalian mitochondrial tRNASer(UCN)
Yokogawa, T., Watanabe, Y., Kumazawa, Y., et al. (1991) · Nucleic Acids Research · doi:10.1093/nar/19.22.6101 · Cited by 73 · see citing works
Impairment of tRNA processing by point mutations in mitochondrial tRNA<sup>Leu(UUR)</sup> associated with mitochondrial diseases
Rossmanith, W., & Karwan, R. M. (1998) · FEBS Letters · doi:10.1016/s0014-5793(98)00928-4 · Cited by 73 · see citing works
Higher-order structure of bovine mitochondrial tRNASerUGA: chemical modification and computer modeling
Watanabe, Y.-I., Kawai, G., Yokogawa, T., et al. (1994) · Nucleic Acids Research · doi:10.1093/nar/22.24.5378 · Cited by 70 · see citing works
A Pathogenesis-associated Mutation in Human Mitochondrial tRNALeu(UUR) Leads to Reduced 3′-End Processing and CCA Addition
Levinger, L., Oestreich, I., Florentz, C., & Mörl, M. (2004) · Journal of Molecular Biology · doi:10.1016/j.jmb.2004.02.008 · Cited by 65 · see citing works
Disease‐related versus polymorphic mutations in human mitochondrial tRNAs
Florentz, C., & Sissler, M. (2001) · EMBO Reports · doi:10.1093/embo-reports/kve111 · Cited by 58 · see citing works
Dual Mode Recognition of Two Isoacceptor tRNAs by Mammalian Mitochondrial Seryl-tRNA Synthetase
Shimada, N., Suzuki, T., & Watanabe, K. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m105150200 · Cited by 56 · see citing works
Nuclear Control of Cloverleaf Structure of Human Mitochondrial tRNALys
Helm, M., & Attardi, G. M. (2004) · Journal of Molecular Biology · doi:10.1016/j.jmb.2004.01.036 · Cited by 53 · see citing works
The 7472insC Mitochondrial DNA Mutation Impairs the Synthesis and Extent of Aminoacylation of tRNASer(UCN) but Not Its Structure or Rate of Turnover
Toompuu, M., Yasukawa, T., Suzuki, T., Hakkinen, T., Spelbrink, J. N., Watanabe, K., & Jacobs, H. T. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m200338200 · Cited by 52 · see citing works
Molecular Phenotype of the np 7472 Deafness-Associated Mitochondrial Mutation in Osteosarcoma Cell Cybrids
Toompuu, M., Tiranti, V., Zeviani, M., & Jacobs, H. T. (1999) · Human Molecular Genetics · doi:10.1093/hmg/8.12.2275 · Cited by 51 · see citing works
The T-loop region of animal mitochondrial tRNASer(AGY) is a main recognition site for homologous seryl-tRNA synthetase
Ueda, T., Yotsumoto, Y., Ikeda, K., et al. (1992) · Nucleic Acids Research · doi:10.1093/nar/20.9.2217 · Cited by 50 · see citing works
Analysis of substrate recognition by the ribonucleoprotein endonuclease RNase P
Christian, E. L., Zahler, N. H., Kaye, N. M., & Harris, M. E. (2002) · Methods · doi:10.1016/s1046-2023(02)00238-4 · Cited by 46 · see citing works
Higher-order structure and thermal instability of bovine mitochondrial tRNASerUGA investigated by proton NMR spectroscopy
Hayashi, I., Kawai, G., & Watanabe, K. (1998) · Journal of Molecular Biology · doi:10.1006/jmbi.1998.2151 · Cited by 37 · see citing works
Mitochondrial tRNALeu isoforms in lung carcinoma cybrid cells containing the np 3243 mtDNA mutation
Meziane, A. E. (1998) · Human Molecular Genetics · doi:10.1093/hmg/7.13.2141 · Cited by 36 · see citing works
Isolation, localization, and physical mapping of a highly polymorphic locus on human chromosome 11q13
Eubanks, J. H., Selleri, L., Hart, R. K., et al. (1991) · Genomics · doi:10.1016/0888-7543(91)90080-x · Cited by 32 · see citing works
Determination of the angle between the anticodon and aminoacyl acceptor stems of yeast phenylalanyl tRNA in solution.
Friederich, M. W., Gast, F. U., Vacano, E., et al. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.11.4803 · Cited by 28 · see citing works
The pathogenic U3271C human mitochondrial tRNALeu(UUR) mutation disrupts a fragile anticodon stem
Wittenhagen, L. M. (2003) · Nucleic Acids Research · doi:10.1093/nar/gkg131 · Cited by 26 · see citing works
The 7472insC mtDNA mutation impairs 5′ and 3′ processing of tRNASer(UCN)
Toompuu, M., Levinger, L., Nadal, A., et al. (2004) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2004.07.181 · Cited by 25 · see citing works
Increased mitochondrial processing intermediates associated with three tRNALeu(UUR) gene mutations
Koga, A., Koga, Y., Akita, Y., Fukiyama, R., Ueki, I., Yatsuga, S., & Matsuishi, T. (2003) · Neuromuscular Disorders · doi:10.1016/s0960-8966(02)00267-5 · Cited by 24 · see citing works
Exercise intolerance, muscle pain and lactic acidaemia associated with a 7497G>A mutation in the tRNASer(UCN) gene
Grafakou, O., Hol, F. A., Schwab, K. O., et al. (2003) · Journal of Inherited Metabolic Disease · doi:10.1023/a:1025960300710 · Cited by 23 · see citing works
Decreased mitochondrial tRNALys steady-state levels and aminoacylation are associated with the pathogenic G8313A mitochondrial DNA mutation
Bacman, S. R., Atencio, D. P., & Moraes, C. T. (2003) · Biochemical Journal · doi:10.1042/bj20030222 · Cited by 21 · see citing works
A T-stem slip in human mitochondrial tRNALeu(CUN) governs its charging capacity
Hao, R. (2005) · Nucleic Acids Research · doi:10.1093/nar/gki677 · Cited by 18 · see citing works
Cytosolic Yeast TrnaHis Is Covalently Modified When Imported into Mitochondria of Trypanosoma brucei
Schneider, A. (1996) · Nucleic Acids Research · doi:10.1093/nar/24.7.1225 · Cited by 11 · see citing works

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Authors: Mollers, M.
Venue: Nucleic Acids Research
Publisher: Oxford University Press (OUP)
Published: 2005-09-25
Volume: 33
Issue: 17
Pages: 5647-5658
ISSN: 0305-1048 (ISSN-L)
1362-4962
DOI: 10.1093/nar/gki876
PMC: PMC1240115
OpenAlex: W2120270701
Cited by: 31
References: 52
Citations by year: 2025: 1 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 1 · 2017: 1 · 2016: 2 · 2015: 4 · 2014: 2 · 2013: 1
FWCI: 1.00
Type: journal article
Language: en
Open access: gold
doi.org
Licence: other-oa
Locations: Nucleic Acids Research (publishedVersion, other-oa)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
DORA Empa (Swiss Federal Laboratories for Materials Science and Technology (Empa)) (submittedVersion, other-oa)
Funders: Deutsche Forschungsgemeinschaft
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
Keywords: mitochondrial tRNA, mitochondrial DNA point mutations, tRNA degradation, mitochondrial translation, respiratory chain deficiency, tRNA structure
MeSH terms: RNA, Mitochondrial, Base Sequence, Cell Line, Child, Child, Preschool, Electron Transport Complex IV, DNA, Mitochondrial, Humans, Male, Molecular Sequence Data, RNA, RNA Precursors, RNA Processing, Post-Transcriptional, RNA, Transfer, Ser, Point Mutation, RNA Stability, Mitochondrial Proteins, Mitochondrial Diseases, Electron Transport Complex I, Aminoacylation

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A new mechanism for mtDNA pathogenesis: impairment of post-transcriptional maturation leads to severe depletion of mitochondrial tRNASer(UCN) caused by T7512C and G7497A point mutations

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