title: A bridge between the aminoacylation and editing domains of leucyl-tRNA synthetase is crucial for its synthetic activity.
abstract: Leucyl-tRNA synthetases (LeuRSs) catalyze the linkage of leucine with tRNA(Leu). LeuRS contains a catalysis domain (aminoacylation) and a CP1 domain (editing). CP1 is inserted 35 Å from the aminoacylation domain. Aminoacylation and editing require CP1 to swing to the coordinated conformation. The neck between the CP1 domain and the aminoacylation domain is defined as the CP1 hairpin. The location of the CP1 hairpin suggests a crucial role in the CP1 swing and domain-domain interaction. Here, the CP1 hairpin of Homo sapiens cytoplasmic LeuRS (hcLeuRS) was deleted or substituted by those from other representative species. Lack of a CP1 hairpin led to complete loss of aminoacylation, amino acid activation, and tRNA binding; however, the mutants retained post-transfer editing. Only the CP1 hairpin from Saccharomyces cerevisiae LeuRS (ScLeuRS) could partly rescue the hcLeuRS functions. Further site-directed mutagenesis indicated that the flexibility of small residues and the charge of polar residues in the CP1 hairpin are crucial for the function of LeuRS.
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Qian
authors: Q
authors: Huang
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Xiao-Long
authors: XL
authors: Zhou
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Qin-Hua
authors: QH
authors: Hu
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Hui-Yan
authors: HY
authors: Lei
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Zhi-Peng
authors: ZP
authors: Fang
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
authors: Peng
authors: P
authors: Yao
authors: Center for RNA Research, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, The Chinese Academy of Sciences, Shanghai 200031, China School of Life Science and Technology, ShanghaiTech University, Shanghai 200031, China.
authors: En-Duo
authors: ED
authors: Wang
mesh_terms: D000595:Amino Acid Sequence; D049148:Aminoacylation / Q000235:genetics*; D002384:Catalysis; D020134:Catalytic Domain / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007935:Leucine-tRNA Ligase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D030781:Organisms, Genetically Modified; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D017393:RNA Editing / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D007935:Leucine-tRNA Ligase
keywords: CP1 hairpin; Homo sapiens cytoplasm; amino acid activation; aminoacylation; leucyl-tRNA synthetase; tRNA binding