Modular pathways for editing non-cognate amino acids by human cytoplasmic leucyl-tRNA synthetase.
To prevent potential errors in protein synthesis, some aminoacyl-transfer RNA (tRNA) synthetases have evolved editing mechanisms to hydrolyze misactivated amino acids (pre-transfer editing) or misacylated tRNAs (post-transfer editing). Class Ia leucyl-tRNA synthetase (LeuRS) may misactivate various natural and non-protein amino acids and then mischarge tRNA(Leu). It is known that the fidelity of prokaryotic LeuRS depends on multiple editing pathways to clear the incorrect intermediates and products in the every step of aminoacylation reaction. Here, we obtained human cytoplasmic LeuRS (hcLeuRS) and tRNA(Leu) (hctRNA(Leu)) with high activity from Escherichia coli overproducing strains to study the synthetic and editing properties of the enzyme. We revealed that hcLeuRS could adjust its editing strategy against different non-cognate amino acids. HcLeuRS edits norvaline predominantly by post-transfer editing; however, it uses mainly pre-transfer editing to edit α-amino butyrate, although both amino acids can be charged to tRNA(Leu). Post-transfer editing as a final checkpoint of the reaction was very important to prevent mis-incorporation in vitro. These results provide insight into the modular editing pathways created to prevent genetic code ambiguity by evolution.
Chen
Xin
X
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences, The Chinese Academy of Sciences, Shanghai 200031, China.
Ma
Jing-Jing
JJ
Tan
Min
M
Yao
Peng
P
Hu
Qing-Hua
QH
Eriani
Gilbert
G
Wang
En-Duo
ED
D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism*; D000613:Aminobutyrates / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007935:Leucine-tRNA Ligase / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D012356:RNA, Transfer, Leu / Q000378:metabolism; D046249:Transfer RNA Aminoacylation; D014633:Valine / Q000031:analogs & derivatives* / Q000378:metabolism
D000613:Aminobutyrates; D012356:RNA, Transfer, Leu; C005313:norvaline; D000604:Amino Acyl-tRNA Synthetases; D007935:Leucine-tRNA Ligase; D014633:Valine
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