Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 20 April 2001:
Vol. 292. no. 5516, pp. 501 - 504
DOI: 10.1126/science.1057718
Prev | Table of Contents | Next

Reports

Enlarging the Amino Acid Set of Escherichia coli by Infiltration of the Valine Coding Pathway

Volker Döring,12 Henning D. Mootz,3 Leslie A. Nangle,4 Tamara L. Hendrickson,4* Valérie de Crécy-Lagard,4 Paul Schimmel,4dagger Philippe Marlière12dagger

Aminoacyl transfer RNA (tRNA) synthetases establish the rules of the genetic code by catalyzing the aminoacylation of tRNAs. For some synthetases, accuracy depends critically on an editing function at a site distinct from the aminoacylation site. Mutants of Escherichia coli that incorrectly charge tRNAVal with cysteine were selected after random mutagenesis of the whole chromosome. All mutations obtained were located in the editing site of valyl-tRNA synthetase. More than 20% of the valine in cellular proteins from such an editing mutant organism could be replaced with the noncanonical aminobutyrate, sterically similar to cysteine. Thus, the editing function may have played a central role in restricting the genetic code to 20 amino acids. Disabling this editing function offers a powerful approach for diversifying the chemical composition of proteins and for emulating evolutionary stages of ambiguous translation.

1 Evologic SA , 4 rue Pierre Fontaine, 91000 Evry, France.
2 UMR8030, Genoscope, 2, rue Gaston Crémieux, CP 5706, 91057 Evry, France.
3 Biochemie/Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Strasse, 35032 Marburg, Germany.
4 The Scripps Research Institute, BCC-379, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
*   Present address: Department of Chemistry, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.

dagger    To whom correspondence should be addressed: E-mail: p.marliere{at}evologic-sa.com or schimmel{at}scripps.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Natural Homolog of tRNA Synthetase Editing Domain Rescues Conditional Lethality Caused by Mistranslation.
Y. E. Chong, X.-L. Yang, and P. Schimmel (2008)
J. Biol. Chem. 283, 30073-30078
   Abstract »    Full Text »    PDF »
Quality control despite mistranslation caused by an ambiguous genetic code.
B. Ruan, S. Palioura, J. Sabina, L. Marvin-Guy, S. Kochhar, R. A. LaRossa, and D. Soll (2008)
PNAS 105, 16502-16507
   Abstract »    Full Text »    PDF »
Unique Residues Crucial for Optimal Editing in Yeast Cytoplasmic Leucyl-tRNA Synthetase Are Revealed by Using a Novel Knockout Yeast Strain.
P. Yao, X.-L. Zhou, R. He, M.-Q. Xue, Y.-G. Zheng, Y.-F. Wang, and E.-D. Wang (2008)
J. Biol. Chem. 283, 22591-22600
   Abstract »    Full Text »    PDF »
Aminoacyl tRNA synthetases and their connections to disease.
S. G. Park, P. Schimmel, and S. Kim (2008)
PNAS 105, 11043-11049
   Abstract »    Full Text »    PDF »
Amino Acid Toxicities of Escherichia coli That Are Prevented by Leucyl-tRNA Synthetase Amino Acid Editing.
V. A. Karkhanis, A. P. Mascarenhas, and S. A. Martinis (2007)
J. Bacteriol. 189, 8765-8768
   Abstract »    Full Text »    PDF »
Phenylalanyl-tRNA synthetase editing defects result in efficient mistranslation of phenylalanine codons as tyrosine.
J. Ling, S. S. Yadavalli, and M. Ibba (2007)
RNA 13, 1881-1886
   Abstract »    Full Text »    PDF »
A present-day aminoacyl-tRNA synthetase with ancestral editing properties.
B. Zhu, M.-W. Zhao, G. Eriani, and E.-D. Wang (2007)
RNA 13, 15-21
   Abstract »    Full Text »    PDF »
Discovery of aminoacyl-tRNA synthetase activity through cell-surface display of noncanonical amino acids.
A. J. Link, M. K. S. Vink, N. J. Agard, J. A. Prescher, C. R. Bertozzi, and D. A. Tirrell (2006)
PNAS 103, 10180-10185
   Abstract »    Full Text »    PDF »
Enzymatic aminoacylation of tRNA with unnatural amino acids.
M. C. T. Hartman, K. Josephson, and J. W. Szostak (2006)
PNAS 103, 4356-4361
   Abstract »    Full Text »    PDF »
Mutational unmasking of a tRNA-dependent pathway for preventing genetic code ambiguity.
A. M. Williams and S. A. Martinis (2006)
PNAS 103, 3586-3591
   Abstract »    Full Text »    PDF »
Binding of misacylated tRNAs to the ribosomal A site.
T. DALE and O. C. UHLENBECK (2005)
RNA 11, 1610-1615
   Abstract »    Full Text »    PDF »
From The Cover: De novo design of defined helical bundles in membrane environments.
B. Bilgicer and K. Kumar (2004)
PNAS 101, 15324-15329
   Abstract »    Full Text »    PDF »
Artificially ambiguous genetic code confers growth yield advantage.
V. Pezo, D. Metzgar, T. L. Hendrickson, W. F. Waas, S. Hazebrouck, V. Doring, P. Marliere, P. Schimmel, and V. de Crecy-Lagard (2004)
PNAS 101, 8593-8597
   Abstract »    Full Text »    PDF »
Cysteinyl-tRNACys Formation in Methanocaldococcus jannaschii: the Mechanism Is Still Unknown.
B. Ruan, H. Nakano, M. Tanaka, J. A. Mills, J. A. DeVito, B. Min, K. B. Low, J. R. Battista, and D. Soll (2004)
J. Bacteriol. 186, 8-14
   Abstract »    Full Text »    PDF »
Crystal structures that suggest late development of genetic code components for differentiating aromatic side chains.
X.-L. Yang, F. J. Otero, R. J. Skene, D. E. McRee, P. Schimmel, and L. Ribas de Pouplana (2003)
PNAS 100, 15376-15380
   Abstract »    Full Text »    PDF »
Nonorthologous replacement of lysyl-tRNA synthetase prevents addition of lysine analogues to the genetic code.
B. C. Jester, J. D. Levengood, H. Roy, M. Ibba, and K. M. Devine (2003)
PNAS 100, 14351-14356
   Abstract »    Full Text »    PDF »
Structure-specific tRNA Determinants for Editing a Mischarged Amino Acid.
K. Beebe, E. Merriman, and P. Schimmel (2003)
J. Biol. Chem. 278, 45056-45061
   Abstract »    Full Text »    PDF »
An Expanded Eukaryotic Genetic Code.
J. W. Chin, T. A. Cropp, J. C. Anderson, M. Mukherji, Z. Zhang, and P. G. Schultz (2003)
Science 301, 964-967
   Abstract »    Full Text »    PDF »
Interstice mutations that block site-to-site translocation of a misactivated amino acid bound to a class I tRNA synthetase.
A. C. Bishop, K. Beebe, and P. R. Schimmel (2003)
PNAS 100, 490-494
   Abstract »    Full Text »    PDF »
Genetic Code Ambiguity. CELL VIABILITY RELATED TO SEVERITY OF EDITING DEFECTS IN MUTANT tRNA SYNTHETASES*.
L. A. Nangle, V. de Crecy Lagard, V. Doring, and P. Schimmel (2002)
J. Biol. Chem. 277, 45729-45733
   Abstract »    Full Text »    PDF »
Cysteine Activation Is an Inherent in Vitro Property of Prolyl-tRNA Synthetases.
I. Ahel, C. Stathopoulos, A. Ambrogelly, A. Sauerwald, H. Toogood, T. Hartsch, and D. Soll (2002)
J. Biol. Chem. 277, 34743-34748
   Abstract »    Full Text »    PDF »
Methanocaldococcus jannaschii Prolyl-tRNA Synthetase Charges tRNAPro with Cysteine.
A. Ambrogelly, I. Ahel, C. Polycarpo, S. Bunjun-Srihari, B. Krett, C. Jacquin-Becker, B. Ruan, C. Kohrer, C. Stathopoulos, U. L. RajBhandary, et al. (2002)
J. Biol. Chem. 277, 34749-34754
   Abstract »    Full Text »    PDF »
An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system.
D. Kiga, K. Sakamoto, K. Kodama, T. Kigawa, T. Matsuda, T. Yabuki, M. Shirouzu, Y. Harada, H. Nakayama, K. Takio, et al. (2002)
PNAS 99, 9715-9720
   Abstract »    Full Text »    PDF »
Plasticity of Recognition of the 3'-End of Mischarged tRNA by Class I Aminoacyl-tRNA Synthetases.
B. E. Nordin and P. Schimmel (2002)
J. Biol. Chem. 277, 20510-20517
   Abstract »    Full Text »    PDF »
Selection and Characterization of Escherichia coli Variants Capable of Growth on an Otherwise Toxic Tryptophan Analogue.
J. M. Bacher and A. D. Ellington (2001)
J. Bacteriol. 183, 5414-5425
   Abstract »    Full Text »    PDF »
Formation of Two Classes of tRNA Synthetases in Relation to Editing Functions and Genetic Code.
P. SCHIMMEL and L. RIBAS DE POUPLANA (2001)
Cold Spring Harb Symp Quant Biol 66, 161-166
   Abstract »    PDF »
Incorporation of azides into recombinant proteins for chemoselective modification by the Staudinger ligation.
K. L. Kiick, E. Saxon, D. A. Tirrell, and C. R. Bertozzi (2002)
PNAS 99, 19-24
   Abstract »    Full Text »    PDF »
Blocking site-to-site translocation of a misactivated amino acid by mutation of a class I tRNA synthetase.
A. C. Bishop, T. K. Nomanbhoy, and P. Schimmel (2002)
PNAS 99, 585-590
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)