title: Modulation of Aminoacylation and Editing Properties of Leucyl-tRNA Synthetase by a Conserved Structural Module.
abstract: A conserved structural module following the KMSKS catalytic loop exhibits α-α-β-α topology in class Ia and Ib aminoacyl-tRNA synthetases. However, the function of this domain has received little attention. Here, we describe the effect this module has on the aminoacylation and editing capacities of leucyl-tRNA synthetases (LeuRSs) by characterizing the key residues from various species. Mutation of highly conserved basic residues on the third α-helix of this domain impairs the affinity of LeuRS for the anticodon stem of tRNA(Leu), which decreases both aminoacylation and editing activities. Two glycine residues on this α-helix contribute to flexibility, leucine activation, and editing of LeuRS from Escherichia coli (EcLeuRS). Acidic residues on the β-strand enhance the editing activity of EcLeuRS and sense the size of the tRNA(Leu) D-loop. Incorporation of these residues stimulates the tRNA-dependent editing activity of the chimeric minimalist enzyme Mycoplasma mobile LeuRS fused to the connective polypeptide 1 editing domain and leucine-specific domain from EcLeuRS. Together, these results reveal the stem contact-fold to be a functional as well as a structural linker between the catalytic site and the tRNA binding domain. Sequence comparison of the EcLeuRS stem contact-fold domain with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS.
authors.affiliation: From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yan
authors.affiliation: From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Ye
authors.affiliation: From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.
authors.forename: Min
authors.initials: M
authors.lastname: Tan
authors.affiliation: the College of Life Sciences, Wuhan University, 299 Bayi Road, Wuhan 430072, Hubei, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: the Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, Institut de Biologie Moléculaire et Cellulaire, 15 Rue René Descartes, Strasbourg 67084, France, and.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Eriani
authors.affiliation: From the State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China, the School of Life Science and Technology, Shanghai Tech University, 319 Yue Yang Road, Shanghai 200031,China, edwang@sibcb.ac.cn.
authors.forename: En-Duo
authors.initials: ED
authors.lastname: Wang
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D002942:Circular Dichroism; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D006801:Humans; D007935:Leucine-tRNA Ligase / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009174:Mycoplasma / Q000201:enzymology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019711:Pyrococcus; D043902:Pyrococcus horikoshii / Q000201:enzymology; D012343:RNA, Transfer / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D000255:Adenosine Triphosphate; D012343:RNA, Transfer; D007935:Leucine-tRNA Ligase
keywords: aminoacyl tRNA Synthetase; aminoacylation; editing; enzyme; evolution; protein synthesis; stem contact fold; transfer RNA (tRNA)
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