title: C-terminal Domain of Leucyl-tRNA Synthetase from Pathogenic Candida albicans Recognizes both tRNASer and tRNALeu.
abstract: Leucyl-tRNA synthetase (LeuRS) is a multidomain enzyme that catalyzes Leu-tRNA(Leu) formation and is classified into bacterial and archaeal/eukaryotic types with significant diversity in the C-terminal domain (CTD). CTDs of both bacterial and archaeal LeuRSs have been reported to recognize tRNA(Leu) through different modes of interaction. In the human pathogen Candida albicans, the cytoplasmic LeuRS (CaLeuRS) is distinguished by its capacity to recognize a uniquely evolved chimeric tRNA(Ser) (CatRNA(Ser)(CAG)) in addition to its cognate CatRNA(Leu), leading to CUG codon reassignment. Our previous study showed that eukaryotic but not archaeal LeuRSs recognize this peculiar tRNA(Ser), suggesting the significance of their highly divergent CTDs in tRNA(Ser) recognition. The results of this study provided the first evidence of the indispensable function of the CTD of eukaryotic LeuRS in recognizing non-cognate CatRNA(Ser) and cognate CatRNA(Leu). Three lysine residues were identified as involved in mediating enzyme-tRNA interaction in the leucylation process: mutation of all three sites totally ablated the leucylation activity. The importance of the three lysine residues was further verified by gel mobility shift assays and complementation of a yeast leuS gene knock-out strain.
authors.affiliation: 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, 200031 Shanghai, China and.
authors.forename: Quan-Quan
authors.initials: QQ
authors.lastname: Ji
authors.affiliation: 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, 200031 Shanghai, China and.
authors.forename: Zhi-Peng
authors.initials: ZP
authors.lastname: Fang
authors.affiliation: 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, 200031 Shanghai, China and.
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, The Chinese Academy of Sciences, 320 Yue Yang Road, 200031 Shanghai, China and.
authors.forename: Zhi-Rong
authors.initials: ZR
authors.lastname: Ruan
authors.affiliation: 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, 200031 Shanghai, China and xlzhou@sibcb.ac.cn.
authors.forename: Xiao-Long
authors.initials: XL
authors.lastname: Zhou
authors.affiliation: 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, 200031 Shanghai, China and School of Life Science and Technology, ShanghaiTech University, 319 Yue Yang Road, 200031 Shanghai, China edwang@sibcb.ac.cn.
authors.forename: En-Duo
authors.initials: ED
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001665:Binding Sites; D002176:Candida albicans / Q000201:enzymology* / Q000378:metabolism; D017124:Conserved Sequence; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055786:Gene Knockout Techniques; D007935:Leucine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D012356:RNA, Transfer, Leu / Q000737:chemistry / Q000378:metabolism*; D012362:RNA, Transfer, Ser / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D005656:Fungal Proteins; D050505:Mutant Proteins; D012331:RNA, Fungal; D012356:RNA, Transfer, Leu; D012362:RNA, Transfer, Ser; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C102042:seryl-tRNA(CAG); D007935:Leucine-tRNA Ligase; D008239:Lysine
keywords: C-terminal domain (carboxyl tail domain, CTD); Candida albicans; aminoacyl-tRNA synthetase; enzyme mechanism; recognition; tRNALeu; tRNASer; transfer RNA (tRNA)