Degenerate connective polypeptide 1 (CP1) domain from human mitochondrial leucyl-tRNA synthetase.
The connective polypeptide 1 (CP1) editing domain of leucyl-tRNA synthetase (LeuRS) from various species either harbors a conserved active site to exclude tRNA mis-charging with noncognate amino acids or is evolutionarily truncated or lost because there is no requirement for high translational fidelity. However, human mitochondrial LeuRS (hmtLeuRS) contains a full-length but degenerate CP1 domain that has mutations in some residues important for post-transfer editing. The significance of such an inactive CP1 domain and a translational accuracy mechanism with different noncognate amino acids are not completely understood. Here, we identified the essential role of the evolutionarily divergent CP1 domain in facilitating hmtLeuRS's catalytic efficiency and endowing enzyme with resistance to AN2690, a broad-spectrum drug acting on LeuRSs. In addition, the canonical core of hmtLeuRS is not stringent for noncognate norvaline (Nva) and valine (Val). hmtLeuRS has a very weak tRNA-independent pre-transfer editing activity for Nva, which is insufficient to remove mis-activated Nva. Moreover, hmtLeuRS chimeras fused with a functional CP1 domain from LeuRSs of other species, regardless of origin, showed restored post-transfer editing activity and acquired fidelity during aminoacylation. This work offers a novel perspective on the role of the CP1 domain in optimizing aminoacylation efficiency.
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and.
Qing
Q
Ye
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and.
Meng
M
Wang
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and.
Zhi-Peng
ZP
Fang
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and.
Zhi-Rong
ZR
Ruan
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and.
Quan-Quan
QQ
Ji
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and xlzhou@sibcb.ac.cn.
Xiao-Long
XL
Zhou
From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai and the School of Life Science and Technology, ShanghaiTech University, 319 Yue Yang Road, Shanghai 200031, China edwang@sibcb.ac.cn.
En-Duo
ED
Wang
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry* / Q000502:physiology; D049148:Aminoacylation; D020134:Catalytic Domain; D002942:Circular Dichroism; D006801:Humans; D006868:Hydrolysis; D007935:Leucine-tRNA Ligase / Q000737:chemistry* / Q000502:physiology; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017393:RNA Editing; D017386:Sequence Homology, Amino Acid; D014633:Valine / Q000031:analogs & derivatives / Q000737:chemistry
D000596:Amino Acids; C005313:norvaline; D000604:Amino Acyl-tRNA Synthetases; C549096:LARS2 protein, human; D007935:Leucine-tRNA Ligase; D014633:Valine
CP1 domain; amino acid; aminoacyl tRNA synthetase; aminoacylation; editing; enzyme catalysis; human mitochondrial leucyl-tRNA synthetase; transfer RNA (tRNA); translation