Aminoacylation and translational quality control strategy employed by leucyl-tRNA synthetase from a human pathogen with genetic code ambiguity

• Published in: Nucleic acids research 2013-11, Vol.41 (21), p.9825-9838
• Main Authors: Zhou, Xiao-Long, Fang, Zhi-Peng, Ruan, Zhi-Rong, Wang, Meng, Liu, Ru-Juan, Tan, Min, Anella, Fabrizio Maria, Wang, En-Duo
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Aminobutyrates - metabolism; Archaea - enzymology; Bacteria - enzymology; Candida albicans - enzymology; Genetic Code; Humans; Leucine-tRNA Ligase - chemistry; Leucine-tRNA Ligase - metabolism; Molecular Sequence Data; Nucleic Acid Enzymes; RNA, Transfer, Leu - metabolism; RNA, Transfer, Ser - metabolism; Sequence Alignment; Species Specificity; Transfer RNA Aminoacylation; Valine - analogs & derivatives; Valine - metabolism
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkt741

Aminoacyl-tRNA synthetases should ensure high accuracy in tRNA aminoacylation. However, the absence of significant structural differences between amino acids always poses a direct challenge for some aminoacyl-tRNA synthetases, such as leucyl-tRNA synthetase (LeuRS), which require editing function to remove mis-activated amino acids. In the cytoplasm of the human pathogen Candida albicans, the CUG codon is translated as both Ser and Leu by a uniquely evolved CatRNA(Ser)(CAG). Its cytoplasmic LeuRS (CaLeuRS) is a crucial component for CUG codon ambiguity and harbors only one CUG codon at position 919. Comparison of the activity of CaLeuRS-Ser(919) and CaLeuRS-Leu(919) revealed yeast LeuRSs have a relaxed tRNA recognition capacity. We also studied the mis-activation and editing of non-cognate amino acids by CaLeuRS. Interestingly, we found that CaLeuRS is naturally deficient in tRNA-dependent pre-transfer editing for non-cognate norvaline while displaying a weak tRNA-dependent pre-transfer editing capacity for non-cognate α-amino butyric acid. We also demonstrated that post-transfer editing of CaLeuRS is not tRNA(Leu) species-specific. In addition, other eukaryotic but not archaeal or bacterial LeuRSs were found to recognize CatRNA(Ser)(CAG). Overall, we systematically studied the aminoacylation and editing properties of CaLeuRS and established a characteristic LeuRS model with naturally deficient tRNA-dependent pre-transfer editing, which increases LeuRS types with unique editing patterns.