title: A Hypertension-Associated tRNAAla Mutation Alters tRNA Metabolism and Mitochondrial Function.
abstract: In this report, we investigated the pathophysiology of a novel hypertension-associated mitochondrial tRNA(Ala) 5655A → G (m.5655A → G) mutation. The destabilization of a highly conserved base pairing (A1-U72) at the aminoacyl acceptor stem by an m.5655A → G mutation altered the tRNA(Ala) function. An in vitro processing analysis showed that the m.5655A → G mutation reduced the efficiency of tRNA(Ala) precursor 5' end cleavage catalyzed by RNase P. By using cybrids constructed by transferring mitochondria from lymphoblastoid cell lines derived from a Chinese family into mitochondrial DNA (mtDNA)-less (ρ(o)) cells, we showed a 41% reduction in the steady-state level of tRNA(Ala) in mutant cybrids. The mutation caused an improperly aminoacylated tRNA(Ala), as suggested by aberrantly aminoacylated tRNA(Ala) and slower electrophoretic mobility of mutated tRNA. A failure in tRNA(Ala) metabolism contributed to variable reductions in six mtDNA-encoded polypeptides in mutant cells, ranging from 21% to 37.5%, with an average of a 29.1% reduction, compared to levels of the controls. The impaired translation caused reduced activities of mitochondrial respiration chains. Furthermore, marked decreases in the levels of mitochondrial ATP and membrane potential were observed in mutant cells. These caused increases in the production of reactive oxygen species in the mutant cybrids. The data provide evidence for the association of the tRNA(Ala) 5655A → G mutation with hypertension.
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Jiang
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: Attardi Institute of Mitochondrial Biomedicine, Wenzhou Medical University, Wenzhou, Zhejiang, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Xue
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Jialing
authors.initials: J
authors.lastname: Yu
authors.affiliation: Attardi Institute of Mitochondrial Biomedicine, Wenzhou Medical University, Wenzhou, Zhejiang, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Attardi Institute of Mitochondrial Biomedicine, Wenzhou Medical University, Wenzhou, Zhejiang, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Yao
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Liu
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Hanqing
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Li
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Institute of Genetics, Zhejiang University, and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou, China Joint Institute of Genetics and Genomic Medicine between Zhejiang University and University of Toronto, Zhejiang University, Hangzhou, Zhejiang, China gminxin88@zju.edu.cn.
authors.forename: Min-Xin
authors.identifier: 0000-0001-5067-6736
authors.initials: MX
authors.lastname: Guan
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044466:Asian People / Q000235:genetics*; D002460:Cell Line; D005260:Female; D006801:Humans; D006973:Hypertension / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000235:genetics / Q000502:physiology*; D009154:Mutation; D009690:Nucleic Acid Conformation; D012313:RNA / Q000235:genetics; D000077278:RNA, Mitochondrial; D012343:RNA, Transfer / Q000378:metabolism*; D012344:RNA, Transfer, Ala / Q000737:chemistry / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000077278:RNA, Mitochondrial; D012344:RNA, Transfer, Ala; D017382:Reactive Oxygen Species; D012313:RNA; D012343:RNA, Transfer