Coexistence of bacterial leucyl-tRNA synthetases with archaeal tRNA binding domains that distinguish tRNA(Leu) in the archaeal mode.
Coexistence of bacterial leucyl-tRNA synthetases with archaeal tRNA binding domains that distinguish tRNA(Leu) in the archaeal mode.
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DOI：
10.1093/nar/gku108
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Wang ED
中科院分区：
文献类型：
--
作者：
Fang ZP;Wang M;Ruan ZR;Tan M;Liu RJ;Zhou M;Zhou XL;Wang ED
亮氨酰-tRNA（transfer RNA）合成酶（LeuRS）是一种多结构域酶，分为细菌型和古/真核型。一般来说，一个特定的LeuRS，其结构域是相同的类型，存在于一个单一的细胞隔室。然而，一些物种，如卤代嗜碱菌Natrialba magadii，编码两种细胞质LeuRS，NmLeuRS 1和NmLeuRS 2，这是天然存在的嵌合酶的第一个例子，具有细菌和古细菌类型的不同结构域。此外，N. magadii编码典型的古细菌tRNAleus。这两个LeuRS的tRNA识别模式，氨酰化和翻译质量控制活动是有趣的问题要解决。在此，活性NmLeuRS 1和NmLeuRS 2在大肠杆菌中基因表达后被成功纯化。在优化的氨酰化条件下，我们发现，他们区分同源NmtRNALeu的古菌模式，而N-末端区域的细菌类型。然而，NmLeuRS 1表现出比NmLeuRS 2高得多的氨酰化和编辑活性，这表明NmLeuRS 1更可能产生Leu-tRNALeu用于蛋白质生物合成。此外，使用NmLeuRS 1作为模型，我们证明了几个非同源氨基酸的误激活，蛋白质合成的准确性主要通过转移后编辑来维持。NmLeuRS/tRNALeu系统的全面研究提供了对氨酰-tRNA合成酶和tRNA的共同进化的详细了解。
Leucyl-tRNA (transfer RNA) synthetase (LeuRS) is a multi-domain enzyme, which is divided into bacterial and archaeal/eukaryotic types. In general, one specific LeuRS, the domains of which are of the same type, exists in a single cell compartment. However, some species, such as the haloalkaliphile Natrialba magadii, encode two cytoplasmic LeuRSs, NmLeuRS1 and NmLeuRS2, which are the first examples of naturally occurring chimeric enzymes with different domains of bacterial and archaeal types. Furthermore, N. magadii encodes typical archaeal tRNALeus. The tRNA recognition mode, aminoacylation and translational quality control activities of these two LeuRSs are interesting questions to be addressed. Herein, active NmLeuRS1 and NmLeuRS2 were successfully purified after gene expression in Escherichia coli. Under the optimized aminoacylation conditions, we discovered that they distinguished cognate NmtRNALeu in the archaeal mode, whereas the N-terminal region was of the bacterial type. However, NmLeuRS1 exhibited much higher aminoacylation and editing activity than NmLeuRS2, suggesting that NmLeuRS1 is more likely to generate Leu-tRNALeu for protein biosynthesis. Moreover, using NmLeuRS1 as a model, we demonstrated misactivation of several non-cognate amino acids, and accuracy of protein synthesis was maintained mainly via post-transfer editing. This comprehensive study of the NmLeuRS/tRNALeu system provides a detailed understanding of the coevolution of aminoacyl-tRNA synthetases and tRNA.
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DOI：
10.1099/00207713-47-3-853
发表时间：
1997-07-01
期刊：
INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY
影响因子：
--
作者：
Kamekura, M;DyallSmith, ML;Kates, M
通讯作者：
Kates, M
影响因子：
64.8
作者：
Hou, SB;Larsen, RW;Alam, M
通讯作者：
Alam, M
影响因子：
11.4
作者：
Cusack, S;Yaremchuk, A;Tukalo, M
通讯作者：
Tukalo, M
影响因子：
16.8
作者：
Fukunaga, R;Yokoyama, S
通讯作者：
Yokoyama, S
影响因子：
14.9
作者：
Du, X;Wang, ED
通讯作者：
Wang, ED