title: Mitochondrial tRNA cleavage by tRNA-targeting ribonuclease causes mitochondrial dysfunction observed in mitochondrial disease.
abstract: Mitochondrial DNA (mtDNA) is a genome possessed by mitochondria. Since reactive oxygen species (ROS) are generated during aerobic respiration in mitochondria, mtDNA is commonly exposed to the risk of DNA damage. Mitochondrial disease is caused by mitochondrial dysfunction, and mutations or deletions on mitochondrial tRNA (mt tRNA) genes are often observed in mtDNA of patients with the disease. Hence, the correlation between mt tRNA activity and mitochondrial dysfunction has been assessed. Then, cybrid cells, which are constructed by the fusion of an enucleated cell harboring altered mtDNA with a ρ(0) cell, have long been used for the analysis due to difficulty in mtDNA manipulation. Here, we propose a new method that involves mt tRNA cleavage by a bacterial tRNA-specific ribonuclease. The ribonuclease tagged with a mitochondrial-targeting sequence (MTS) was successfully translocated to the mitochondrial matrix. Additionally, mt tRNA cleavage, which resulted in the decrease of cytochrome c oxidase (COX) activity, was observed.
authors: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: atetsu@mail.ecc.u-tokyo.ac.jp.
authors: Tetsuhiro
authors: T
authors: Ogawa
authors: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors: Ayano
authors: A
authors: Shimizu
authors: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors: Kazutoshi
authors: K
authors: Takahashi
authors: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors: Makoto
authors: M
authors: Hidaka
authors: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: amasaki@mail.ecc.u-tokyo.ac.jp.
authors: Haruhiko
authors: H
authors: Masaki
mesh_terms: D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000235:genetics; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D028361:Mitochondrial Diseases / Q000235:genetics; D008969:Molecular Sequence Data; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D000077278:RNA, Mitochondrial; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004272:DNA, Mitochondrial; D029968:Escherichia coli Proteins; D000077278:RNA, Mitochondrial; D011994:Recombinant Proteins; C501879:colicin D immunity protein, E coli; D049452:Green Fluorescent Proteins; D012313:RNA; D012343:RNA, Transfer; C477826:COX8C protein, human; D003576:Electron Transport Complex IV
keywords: Mitochondrial disease; Mitochondrial tRNA; Ribonuclease