Journal
NUCLEIC ACIDS RESEARCH
Volume 37, Issue 20, Pages 6881-6895Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkp697
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Funding
- Centre National de la Recherche Scientifique (CNRS)
- Universite de Strasbourg, Association Francaise contre les Myopathies
- Action Concertee Incitative (ACI BCMS)
- Ministry of Education, Science, Sports, and Culture of Japan
- JSPS
- New Energy and Industrial Technology Development Organization (NEDO)
- Ministere de l'Enseignement Superieur et de la Recherche
- AFM
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Primary and secondary structures of mammalian mitochondrial (mt) tRNAs are divergent from canonical tRNA structures due to highly skewed nucleotide content and large size variability of D- and T-loops. The nonconservation of nucleotides involved in the expected network of tertiary interactions calls into question the rules governing a functional L-shaped three-dimensional (3D) structure. Here, we report the solution structure of human mt-tRNA(Asp) in its native post-transcriptionally modified form and as an in vitro transcript. Probing performed with nuclease S1, ribonuclease V1, dimethylsulfate, diethylpyrocarbonate and lead, revealed several secondary structures for the in vitro transcribed mt-tRNA(Asp) including predominantly the cloverleaf. On the contrary, the native tRNA(Asp) folds into a single cloverleaf structure, highlighting the contribution of the four newly identified post-transcriptional modifications to correct folding. Reactivities of nucleotides and phosphodiester bonds in the native tRNA favor existence of a full set of six classical tertiary interactions between the D-domain and the variable region, forming the core of the 3D structure. Reactivities of D- and T-loop nucleotides support an absence of interactions between these domains. According to multiple sequence alignments and search for conservation of Leontis-Westhof interactions, the tertiary network core building rules apply to all tRNA(Asp) from mammalian mitochondria.
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