Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability

Min Gang Xu; Juan Li; Xing Du; En Duo Wang

doi:10.1016/j.bbrc.2004.03.180

Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability

Min Gang Xu, Juan Li, Xing Du, En Duo Wang

Research output: Contribution to journal › Article › peer-review

29 Scopus citations

Abstract

Leucyl-tRNA synthetase (LeuRS) catalyzes the leucylation of tRNA ^Leu. To maintain the fidelity of protein biosynthesis, LeuRS also catalyzes the editing reaction. In the present work, highly conserved T252 in the T-rich region within CP1 domain of Escherichia coli LeuRS was mutated to G, D, or E. Steady-state kinetic of aminoacylation, and combined editing assays indicated that not only the size of the amino acid but also the absence of hydrogen bonds between T252 and adjacent molecules may affect the editing. It is further confirmed by in vivo experiments using the temperature-sensitive strain KL231 (ΔleuS), which revealed the arrested growth of bacterial cells bearing mutants with highly impaired editing activity in the presence of leucine analog.

Original language	English
Pages (from-to)	11-16
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	318
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2004.03.180
State	Published - 21 May 2004
Externally published	Yes

Keywords

Aminoacyl-tRNA synthetase
Editing
Temperature-sensitive strain

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10.1016/j.bbrc.2004.03.180

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title = "Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability",

abstract = "Leucyl-tRNA synthetase (LeuRS) catalyzes the leucylation of tRNA Leu. To maintain the fidelity of protein biosynthesis, LeuRS also catalyzes the editing reaction. In the present work, highly conserved T252 in the T-rich region within CP1 domain of Escherichia coli LeuRS was mutated to G, D, or E. Steady-state kinetic of aminoacylation, and combined editing assays indicated that not only the size of the amino acid but also the absence of hydrogen bonds between T252 and adjacent molecules may affect the editing. It is further confirmed by in vivo experiments using the temperature-sensitive strain KL231 (ΔleuS), which revealed the arrested growth of bacterial cells bearing mutants with highly impaired editing activity in the presence of leucine analog.",

keywords = "Aminoacyl-tRNA synthetase, Editing, Temperature-sensitive strain",

author = "Xu, {Min Gang} and Juan Li and Xing Du and Wang, {En Duo}",

note = "Funding Information: This work was funded by the Chinese Academy of Sciences (Grant KSCX2-2-04), 863 project of China (Grant 2001AA235071) and Shanghai Committee of Science and Technology (Grant 02DJ140567). We are grateful to Professor You-Shang Zhang for carefully reading the manuscript.",

year = "2004",

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doi = "10.1016/j.bbrc.2004.03.180",

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journal = "Biochemical and Biophysical Research Communications",

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AU - Xu, Min Gang

AU - Li, Juan

AU - Du, Xing

AU - Wang, En Duo

N1 - Funding Information: This work was funded by the Chinese Academy of Sciences (Grant KSCX2-2-04), 863 project of China (Grant 2001AA235071) and Shanghai Committee of Science and Technology (Grant 02DJ140567). We are grateful to Professor You-Shang Zhang for carefully reading the manuscript.

PY - 2004/5/21

Y1 - 2004/5/21

N2 - Leucyl-tRNA synthetase (LeuRS) catalyzes the leucylation of tRNA Leu. To maintain the fidelity of protein biosynthesis, LeuRS also catalyzes the editing reaction. In the present work, highly conserved T252 in the T-rich region within CP1 domain of Escherichia coli LeuRS was mutated to G, D, or E. Steady-state kinetic of aminoacylation, and combined editing assays indicated that not only the size of the amino acid but also the absence of hydrogen bonds between T252 and adjacent molecules may affect the editing. It is further confirmed by in vivo experiments using the temperature-sensitive strain KL231 (ΔleuS), which revealed the arrested growth of bacterial cells bearing mutants with highly impaired editing activity in the presence of leucine analog.

AB - Leucyl-tRNA synthetase (LeuRS) catalyzes the leucylation of tRNA Leu. To maintain the fidelity of protein biosynthesis, LeuRS also catalyzes the editing reaction. In the present work, highly conserved T252 in the T-rich region within CP1 domain of Escherichia coli LeuRS was mutated to G, D, or E. Steady-state kinetic of aminoacylation, and combined editing assays indicated that not only the size of the amino acid but also the absence of hydrogen bonds between T252 and adjacent molecules may affect the editing. It is further confirmed by in vivo experiments using the temperature-sensitive strain KL231 (ΔleuS), which revealed the arrested growth of bacterial cells bearing mutants with highly impaired editing activity in the presence of leucine analog.

KW - Aminoacyl-tRNA synthetase

KW - Editing

KW - Temperature-sensitive strain

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VL - 318

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 1

ER -

Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability

Abstract

Keywords

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