Search for differences in post-transcriptional modification patterns of mitochondrial DNA-encoded wild-type and mutant human tRNALys and tRNALeu(UUR)
| Autor: | Anne Chomyn, Catherine Florentz, Giuseppe Attardi, Mark Helm |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Mitochondrial DNA
RNA Transfer Leu Molecular Sequence Data Mutant Biology Mitochondrion DNA Mitochondrial Cell Line Mitochondrial myopathy MELAS Syndrome Genetics medicine Humans Base Sequence Wild type RNA medicine.disease Molecular biology MERRF Syndrome Post-transcriptional modification Biochemistry Mutation Transfer RNA Autoradiography Nucleic Acid Conformation RNA Transfer Lys Chromatography Thin Layer Protein Processing Post-Translational Caltech Library Services Research Article |
| Zdroj: | Nucleic Acids Research. 27:756-763 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/27.3.756 |
| Popis: | Post-transcriptional modifications are characteristic features of tRNAs and have been shown in a number of cases to influence both their structural and functional properties, including structure stabilization, amino-acylation and codon recognition. We have developed an approach which allows the investigation of the post-transcriptional modification patterns of human mitochondrial wild-type and mutant tRNAs at both the qualitative and the quantitative levels. Specific tRNA species are long-term labeled in vivo with [P-32]orthophosphate, isolated in a highly selective way, enzymatically digested to mononucleotides and then subjected to two-dimensional thin layer chromatographic analysis. The wild-type tRNA(LyS) and the corresponding tRNALyS carrying the A8344G mutation associated with the MERRF (Myoclonic Epilepsy with Ragged Red Fibers) syndrome exhibit the same modified nucleotides at the same molar concentrations. By contrast, a quantitatively different modification pattern was observed between the wild-type tRNA(LeU(UUR)) and its counterpart carrying the A3243G mutation associated with the MELAS (Mitochondrial Myopathy, Encephalopathy with Lactic Acidosis and Stroke-like episodes) syndrome, the latter exhibiting a 50% decrease in m(2)G content. Complementary sequencing of tRNA(Leu(UUR)) has allowed the localization of this modification at position 10 within the D-stem of the tRNA. The decreased lever of this modification may have important implications for understanding the molecular mechanism underlying the MELAS-associated mitochondrial dysfunction. |
| Databáze: | OpenAIRE |
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