Unique residues crucial for optimal editing in yeast cytoplasmic leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain

	Unique residues crucial for optimal editing in yeast cytoplasmic leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain
	Yao, P; Zhou, XL; He, R; Xue, MQ; Zheng, YG; Wang, YF; Wang, ED
刊名	JOURNAL OF BIOLOGICAL CHEMISTRY
	2008
卷号	283 期号:33 页码:22591-22600
通讯作者	Wang, ED (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Grad Sch,State Key Lab Mol Biol, Inst Biochem & Cell Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,edwang@sibs.ac.cn
英文摘要	Leucyl-tRNA synthetase (LeuRS) contains an editing domain that discriminates leucine from noncognate amino acids to ensure translational fidelity. In this study, a knock-out strain for Saccharomyces cerevisiae LeuRS was constructed to analyze in vivo the tRNA aminoacylation properties of S. cerevisiae and human cytoplasmic LeuRSs. The activities of several editing-defective mutants of ycLeuRS were determined in vitro and compared with those obtained in vivo in a complementation assay performed in the knock-out strain. The editing activities of these mutants were analyzed in the presence of either norvaline, a leucine analogue, or AN2690, a specific inhibitor that targets the editing active site. In general, the in vivo data are consistent with those obtained in vitro. Our results show that ycLeuRS post-transfer editing plays a crucial role in the establishment of the aminoacylation fidelity. When impaired, the viability of cells bearing editing-defective mutants is drastically decreased in the presence of noncognate amino acid. This study also emphasizes the crucial function of some semi-conserved residues around the editing site in modulating the editing efficiency. The assay system can be used to test the effect of compounds that potentially target the aminoacylation or editing active site of fungal LeuRS.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	IN-VIVO SELECTION ; ESCHERICHIA-COLI ; SACCHAROMYCES-CEREVISIAE ; CELL VIABILITY ; GENETIC-CODE ; AQUIFEX-AEOLICUS ; AMINO-ACIDS ; MITOCHONDRIAL ; RECOGNITION ; TRNA(LEU)
收录类别	SCI
语种	英语
WOS记录号	WOS:000258321000028
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1478]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Yao, P,Zhou, XL,He, R,et al. Unique residues crucial for optimal editing in yeast cytoplasmic leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2008,283(33):22591-22600.
APA	Yao, P.,Zhou, XL.,He, R.,Xue, MQ.,Zheng, YG.,...&Wang, ED.(2008).Unique residues crucial for optimal editing in yeast cytoplasmic leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain.JOURNAL OF BIOLOGICAL CHEMISTRY,283(33),22591-22600.
MLA	Yao, P,et al."Unique residues crucial for optimal editing in yeast cytoplasmic leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain".JOURNAL OF BIOLOGICAL CHEMISTRY 283.33(2008):22591-22600.