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Decreased aminoacylation in pathology-related mutants of mitochondrial tRNATyr is associated with structural perturbations in tRNA architecture
- Source :
- RNA, RNA, Cold Spring Harbor Laboratory Press, 2008, 14 (4), pp.641-8. ⟨10.1261/rna.938108⟩
- Publication Year :
- 2008
- Publisher :
- HAL CCSD, 2008.
-
Abstract
- A growing number of human pathologies are ascribed to mutations in mitochondrial tRNA genes. Here, we report biochemical investigations on three mt-tRNATyr molecules with point substitutions associated with diseases. The mutations occur in the atypical T- and D-loops at positions homologous to those involved in the tertiary interaction network of canonical tRNAs. They do not correspond to tyrosine identity positions and likely do not contact the mitochondrial tyrosyl-tRNA synthetase during the aminoacylation process. The impact of these substitutions on mt-tRNATyr tyrosylation and structure was investigated using the corresponding tRNA transcripts. In vitro tyrosylation efficiency is decreased 600-fold for mutant A22G (mitochondrial gene mutation T5874C), 40-fold for G15A (C5877T), and is without significant effect on U54C (A5843G). Comparative solution probings with lead and nucleases on mutant and wild-type tRNATyr molecules reveal a greater sensitivity to single-strand specific probes for mutants G15A and A22G. For both transcripts, the mutation triggers a structural destabilization in the D-loop that propagates toward the anticodon arm and thus hinders efficient tyrosylation. Further probing analysis combined with phylogenetic data support the participation of G15 and A22 in the tertiary network of human mt-tRNATyr via nonclassical Watson–Crick G15–C48 and G13–A22 pairings. In contrast, the pathogenic effect of the tyrosylable mutant U54C, where structure is only marginally affected, has to be sought at another level of the tRNATyr life cycle.
- Subjects :
- Models, Molecular
Mitochondrial Diseases
RNA, Mitochondrial
RNA Stability
Mutant
Mitochondrion
MESH: Base Sequence
medicine.disease_cause
0302 clinical medicine
Transfer RNA Aminoacylation
Genetics
0303 health sciences
Mutation
MESH: Mitochondrial Diseases
MESH: RNA Stability
tRNATyr
mitochondria
RNA, Transfer, Tyr
MESH: Nucleic Acid Conformation
Transfer RNA
MESH: Models, Molecular
tyrosyl-tRNA synthetase
Molecular Sequence Data
Aminoacylation
[SDV.BC]Life Sciences [q-bio]/Cellular Biology
In Vitro Techniques
Biology
03 medical and health sciences
MESH: RNA
Report
medicine
MESH: RNA, Transfer, Tyr
Humans
Point Mutation
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Molecular Biology
030304 developmental biology
MESH: Point Mutation
aminoacylation
disease
MESH: Humans
MESH: Molecular Sequence Data
Base Sequence
Point mutation
RNA
structure probing
MESH: Transfer RNA Aminoacylation
Nucleic Acid Conformation
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 13558382 and 14699001
- Database :
- OpenAIRE
- Journal :
- RNA, RNA, Cold Spring Harbor Laboratory Press, 2008, 14 (4), pp.641-8. ⟨10.1261/rna.938108⟩
- Accession number :
- edsair.doi.dedup.....975511c262a8f4314ad6c411e7b63293