1. A deafness-associated tRNA mutation caused pleiotropic effects on the m1G37 modification, processing, stability and aminoacylation of tRNAIle and mitochondrial translation
- Author
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Min-Xin Guan, Mi Zhou, Meng Wang, Feilong Meng, Xiaoting Mao, Yong Fu, Zhenzhen Ye, Jing Zheng, Yun Xiao, Jiaxi Lin, Tianxiang Lin, and Xiaohui Cang
- Subjects
Male ,AcademicSubjects/SCI00010 ,Mitochondrial translation ,Mitochondrion ,Oxidative Phosphorylation ,Adenosine Triphosphate ,0302 clinical medicine ,Ethnicity ,RNA Processing, Post-Transcriptional ,Transfer RNA Aminoacylation ,RNA, Transfer, Ile ,Membrane Potential, Mitochondrial ,tRNA Methyltransferases ,0303 health sciences ,Genetic Pleiotropy ,Middle Aged ,Recombinant Proteins ,Mitochondria ,Pedigree ,Cell biology ,Methanocaldococcus ,Transfer RNA ,Female ,Maternal Inheritance ,Adult ,RNase P ,Archaeal Proteins ,Hearing Loss, Sensorineural ,Aminoacylation ,Molecular Dynamics Simulation ,Biology ,DNA, Mitochondrial ,Methylation ,Cell Line ,Young Adult ,03 medical and health sciences ,RNA and RNA-protein complexes ,Autophagy ,Genetics ,Humans ,Point Mutation ,Isoleucine ,030304 developmental biology ,Base Sequence ,Point mutation ,TRNA Methyltransferase ,Protein Biosynthesis ,Nucleic Acid Conformation ,030217 neurology & neurosurgery - Abstract
Defects in the posttranscriptional modifications of mitochondrial tRNAs have been linked to human diseases, but their pathophysiology remains elusive. In this report, we investigated the molecular mechanism underlying a deafness-associated tRNAIle 4295A>G mutation affecting a highly conserved adenosine at position 37, 3′ adjacent to the tRNA’s anticodon. Primer extension and methylation activity assays revealed that the m.4295A>G mutation introduced a tRNA methyltransferase 5 (TRMT5)-catalyzed m1G37 modification of tRNAIle. Molecular dynamics simulations suggested that the m.4295A>G mutation affected tRNAIle structure and function, supported by increased melting temperature, conformational changes and instability of mutated tRNA. An in vitro processing experiment revealed that the m.4295A>G mutation reduced the 5′ end processing efficiency of tRNAIle precursors, catalyzed by RNase P. We demonstrated that cybrid cell lines carrying the m.4295A>G mutation exhibited significant alterations in aminoacylation and steady-state levels of tRNAIle. The aberrant tRNA metabolism resulted in the impairment of mitochondrial translation, respiratory deficiency, decreasing membrane potentials and ATP production, increasing production of reactive oxygen species and promoting autophagy. These demonstrated the pleiotropic effects of m.4295A>G mutation on tRNAIle and mitochondrial functions. Our findings highlighted the essential role of deficient posttranscriptional modifications in the structure and function of tRNA and their pathogenic consequence of deafness.
- Published
- 2021