近期Nucleic Acids Research在线发表了上海生化与细胞所王恩多研究组的研究论文：人细胞质亮氨酰-tRNA合成酶编校非对应氨基酸的模块式途径。
为了防止潜在的蛋白质错误合成，某些氨基酰-tRNA合成酶具有编校功能去除错误活化的氨基酸或者错误接载的氨基酰化tRNA，通过控制蛋白质的生物合成原料的质量，对蛋白质合成进行质量控制。该实验室已经证明亮氨酰-tRNA合成酶(LeuRS)能够通过不依赖-或依赖tRNA转移前编校途径水解误活化的非对应氨基酸，即使误活化的氨基酸转移到tRNA分子上也可以通过转移后编校途径将其水解。 LeuRS通过多条编校途径在反应的各个步骤清除错误的反应产物和中间物。
王恩多研究组的博士研究生陈鑫和马晶晶成功地首次在大肠杆菌基因表达体系中得到了高活力的人胞质亮氨酰-tRNA合成酶(hcLeuRS)和亮氨酸tRNA，建立了一个高效的测定hcLeuRS合成和编校活力方法和体外研究体系。发现hcLeuRS对不同的非对应氨基酸的胁迫采取不同的编校策略对产物进行质量控制，虽然亮氨酸的类似物正缬氨酸(Nva)和2-氨基丁酸(ABA)都能被错误地载入tRNA分子的3’端，但主要通过转移后的编校途径去除Nva，去除ABA则偏爱转移前的编校途径。转移后编校作为tRNA氨基酰化最后的一个质量控制的检查点对于避免氨基酸的错误掺入发挥着至关重要的作用。该研究结果将有助于更加深入、全面地认识生物进化过程中产生的模块式编校途径在保持遗传信息由RNA传递到蛋白质的精确性发挥着重要作用。
该研究得到中国科学院，科技部重大科学研究计划，国家自然科学基金委，上海市科委的资助。
生物谷推荐英文摘要：
Nucleic Acids Research, doi:10.1093/nar/gkq763
Modular pathways for editing non-cognate amino acids by human cytoplasmic leucyl-tRNA synthetase
Xin Chen1, Jing-Jing Ma1, Min Tan1, Peng Yao1, Qing-Hua Hu1, Gilbert Eriani2 and En-Duo Wang1,*
1State 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, 320 Yue Yang Road, Shanghai 200031, China and 2Architecture et Réactivité de l’ARN, Institut de Biologie Moléculaire et Cellulaire du CNRS, Université de Strasbourg, 15 rue René Descartes 67084 Strasbourg, France
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 tRNALeu. 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 tRNALeu (hctRNALeu) 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 -amino butyrate, although both amino acids can be charged to tRNALeu. 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.
