In vivo identification of essential nucleotides in tRNA(Leu) to its functions by using a constructed yeast tRNA(Leu) knockout strain

	In vivo identification of essential nucleotides in tRNA(Leu) to its functions by using a constructed yeast tRNA(Leu) knockout strain
	Huang, Q; Yao, P; Eriani, G; Wang, ED
刊名	NUCLEIC ACIDS RESEARCH
	2012
卷号	40 期号:20 页码:10463-10477
通讯作者	Wang, ED (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, State Key Lab Mol Biol, Inst Biochem & Cell Biol,Ctr RNA Res, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,edwang@sibs.ac.cn
英文摘要	The fidelity of protein biosynthesis requires the aminoacylation of tRNA with its cognate amino acid catalyzed by aminoacyl-tRNA synthetase with high levels of accuracy and efficiency. Crucial bases in tRNA(Leu) to aminoacylation or editing functions of leucyl-tRNA synthetase have been extensively studied mainly by in vitro methods. In the present study, we constructed two Saccharomyces cerevisiae tRNA(Leu) knockout strains carrying deletions of the genes for tRNA(Leu)(GAG) and tRNA(Leu)(UAG). Disrupting the single gene encoding tRNA(Leu)(GAG) had no phenotypic consequence when compared to the wild-type strain. While disrupting the three genes for tRNA(Leu)(UAG) had a lethal effect on the yeast strain, indicating that tRNA(Leu)(UAG) decoding capacity could not be compensated by another tRNA(Leu) isoacceptor. Using the triple tRNA knockout strain and a randomly mutated library of tRNA(Leu)(UAG), a selection to identify critical tRNA(Leu) elements was performed. In this way, mutations inducing in vivo decreases of tRNA levels or aminoacylation or editing ability by leucyl-tRNA synthetase were identified. Overall, the data showed that the triple tRNA knockout strain is a suitable tool for in vivo studies and identification of essential nucleotides of the tRNA.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	TRANSFER-RNA SYNTHETASE ; SACCHAROMYCES-CEREVISIAE ; ESCHERICHIA-COLI ; QUALITY-CONTROL ; ANTICODON LOOP ; AMINOACYLATION ; GENE ; TRANSCRIPTION ; MUTATIONS ; ACID
收录类别	SCI
语种	英语
WOS记录号	WOS:000310970700051
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/684]
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	Huang, Q,Yao, P,Eriani, G,et al. In vivo identification of essential nucleotides in tRNA(Leu) to its functions by using a constructed yeast tRNA(Leu) knockout strain[J]. NUCLEIC ACIDS RESEARCH,2012,40(20):10463-10477.
APA	Huang, Q,Yao, P,Eriani, G,&Wang, ED.(2012).In vivo identification of essential nucleotides in tRNA(Leu) to its functions by using a constructed yeast tRNA(Leu) knockout strain.NUCLEIC ACIDS RESEARCH,40(20),10463-10477.
MLA	Huang, Q,et al."In vivo identification of essential nucleotides in tRNA(Leu) to its functions by using a constructed yeast tRNA(Leu) knockout strain".NUCLEIC ACIDS RESEARCH 40.20(2012):10463-10477.