A Human Disease-causing Point Mutation in Mitochondrial Threonyl-tRNA Synthetase Induces Both Structural and Functional Defects

doi:10.1074/jbc.M115.700849

	A Human Disease-causing Point Mutation in Mitochondrial Threonyl-tRNA Synthetase Induces Both Structural and Functional Defects
	Wang, Yong1,2 ; Zhou, Xiao-Long 1; Ruan, Zhi-Rong 1; Liu, Ru-Juan 1; Eriani, Gilbert 3; Wang, En-Duo1,2
	2016-03-18
发表期刊	JOURNAL OF BIOLOGICAL CHEMISTRY (IF:4.0[JCR-2023],4.4[5-Year])
ISSN	1083351X
卷号	291 期号:12 页码:6507-6520
发表状态	已发表
DOI	10.1074/jbc.M115.700849
摘要	Mitochondria require all translational components, including aminoacyl-tRNA synthetases (aaRSs), to complete organelle protein synthesis. Some aaRS mutations cause mitochondrial disorders, including human mitochondrial threonyl-tRNA synthetase (hmtThrRS) (encoded by TARS2), the P282L mutation of which causes mitochondrial encephalomyopathies. However, its catalytic and structural consequences remain unclear. Herein, we cloned TARS2 and purified the wild-type and P282L mutant hmtThrRS. hmtThrRS misactivates non-cognate Ser and uses post-transfer editing to clear erroneously synthesized products. In vitro and in vivo analyses revealed that the mutation induces a decrease in Thr activation, aminoacylation, and proofreading activities and a change in the protein structure and/or stability, which might cause reduced catalytic efficiency. We also identified a splicing variant of TARS2 mRNA lacking exons 8 and 9, the protein product of which is targeted into mitochondria. In HEK293T cells, the variant does not dimerize and cannot complement the ThrRS knock-out strain in yeast, suggesting that the truncated protein is inactive and might have a non-canonical function, as observed for other aaRS fragments. The present study describes the aminoacylation and editing properties of hmtThrRS, clarifies the molecular consequences of the P282L mutation, and shows that the yeast ThrRS-deletion model is suitable to test pathology-associated point mutations or alternative splicing variants of mammalian aaRS mRNAs.
关键词	alternative splicing aminoacyl-tRNA synthetase enzyme kinetics mitochondria mitochondrial disease threonyl-tRNA synthetase
收录类别	SCI ; EI
语种	英语
资助项目	Committee of Science and Technology in Shanghai[12JC1409700] ; Committee of Science and Technology in Shanghai[15ZR1446500]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000372894200037
出版者	AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
EI入藏号	20161902345054
EI主题词	Amino acids ; Biosynthesis ; Cloning ; Mammals ; Proteins ; Yeast
EI分类号	Bioengineering and Biology:461 ; Organic Compounds:804.1 ; Food Products:822.3
WOS关键词	EDITING ACTIVITY ; QUALITY-CONTROL ; PROTEIN ; DOMAIN ; CODON ; MISTRANSLATION ; TRANSLATION ; MECHANISM ; ERRORS
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1892
专题	生命科学与技术学院生命科学与技术学院_特聘教授组_王恩多组生命科学与技术学院_博士生
通讯作者	Zhou, Xiao-Long; Wang, En-Duo
作者单位	1.Univ Chinese Acad Sci, Shanghai Inst Biol Sci, State Key Lab Mol Biol,Inst Biochem & Cell Biol, Chinese Acad Sci,Ctr Excellence Mol Cell Sci, Shanghai 200031, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, 319 Yue Yang Rd, Shanghai 200031, Peoples R China 3.Univ Strasbourg, Inst Biol Mol & Cellulaire, CNRS, Architecture & React ARN,UPR9002, 15 Rue Rene Descartes, F-67084 Strasbourg, France
第一作者单位	生命科学与技术学院
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Wang, Yong,Zhou, Xiao-Long,Ruan, Zhi-Rong,et al. A Human Disease-causing Point Mutation in Mitochondrial Threonyl-tRNA Synthetase Induces Both Structural and Functional Defects[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2016,291(12):6507-6520.
APA	Wang, Yong,Zhou, Xiao-Long,Ruan, Zhi-Rong,Liu, Ru-Juan,Eriani, Gilbert,&Wang, En-Duo.(2016).A Human Disease-causing Point Mutation in Mitochondrial Threonyl-tRNA Synthetase Induces Both Structural and Functional Defects.JOURNAL OF BIOLOGICAL CHEMISTRY,291(12),6507-6520.
MLA	Wang, Yong,et al."A Human Disease-causing Point Mutation in Mitochondrial Threonyl-tRNA Synthetase Induces Both Structural and Functional Defects".JOURNAL OF BIOLOGICAL CHEMISTRY 291.12(2016):6507-6520.