HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 40, 41960-41965, October 1, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/40/41960    most recent M401937200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jia, J. Articles by Jin, Y.-X. Search for Related Content PubMed PubMed Citation Articles by Jia, J. Articles by Jin, Y.-X. Social Bookmarking                      What's this?

Residues Lys-149 and Glu-153 Switch the Aminoacylation of tRNA^Trp in Bacillus subtilis^*

Jie Jia, Xiang-Long Chen, Li-Tao Guo, Ya-Dong Yu¶, Jian-Ping Ding¶, and You-Xin Jin||

From the State Key Laboratory of Molecular Biology, ^¶Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Science, Shanghai 200031, China

Tryptophanyl-tRNA synthetase (TrpRS) consists of two identical subunits that induce the cross-subunit binding mode of tRNA^Trp. It has been shown that eubacterial and eukaryotic TrpRSs cannot efficiently cross-aminoacylate the corresponding tRNA^Trp. Although the identity elements in tRNA^Trp that confer the species-specific recognition have been identified, the corresponding elements in TrpRS have not yet been reported. In this study two residues, Lys-149 and Glu-153, were identified as being crucial for the accurate recognition of tRNA^Trp. These residues reside adjacent to the binding pocket for Trp-AMP and show phylogenic diversities in the charge on their side chains between eubacteria and eukaryotes. Single mutagenesis at Lys-149 or Glu-153 reduced the activity of TrpRS in the activation of Trp. The reduction was less than that caused by the double mutant WBHA (K149D/E153R). It is unusual that E153G had no detectable activity in the activation of Trp unless tRNA^Trp was added to the reaction. In addition, we successfully switched the species specificity of Bacillus subtilis TrpRS recognition of tRNA^Trp. The affinity of WBHA, K149E and E153K to human tRNA^Trp was 31-, 13.5-, and 12.9-fold greater than that of wild type B. subtilis TrpRS, respectively. Indeed WBHA and E153K were found to prefer genuine human tRNA^Trp to their cognate eubacteria tRNA^Trp.

Received for publication, February 22, 2004 , and in revised form, July 7, 2004.

^* This work is supported by Grant 2002CB512803 from State Key Programs Basic Research of China, Grant 30370325 from the National Natural Science Foundation of China, and Grant 02DJ14004 from the Shanghai Council of Science and Technology. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

These authors contributed equally to this work.

^|| To whom correspondence should be addressed: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Science, Yueyang Road No. 320, Shanghai 200031, China. Tel.: 86-21-5492-1122; Fax: 86-21-5492-1011; E-mail: yxjin{at}sunm.shcnc.ac.cn.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L.-T. Guo, X.-L. Chen, B.-T. Zhao, Y. Shi, W. Li, H. Xue, and Y.-X. Jin Human tryptophanyl-tRNA synthetase is switched to a tRNA-dependent mode for tryptophan activation by mutations at V85 and I311 Nucleic Acids Res., September 27, 2007; 35(17): 5934 - 5943. [Abstract] [Full Text] [PDF]

				N. Shen, L. Guo, B. Yang, Y. Jin, and J. Ding Structure of human tryptophanyl-tRNA synthetase in complex with tRNATrp reveals the molecular basis of tRNA recognition and specificity Nucleic Acids Res., June 23, 2006; 34(11): 3246 - 3258. [Abstract] [Full Text] [PDF]