Fidelity escape by the unnatural amino acid β-hydroxynorvaline: an efficient substrate for Escherichia coli threonyl-tRNA synthetase with toxic effects on growth

Biochemistry
Anand MinajigiChristopher S Francklyn

Abstract

In all living systems, the fidelity of translation is maintained in part by the editing mechanisms of aminoacyl-tRNA synthetases (ARSs). Some nonproteogenic amino acids, including β-hydroxynorvaline (HNV) are nevertheless efficiently aminoacylated and become incorporated into proteins. To investigate the basis of HNV's ability to function in protein synthesis, the utilization of HNV by Escherichia coli threonyl-tRNA synthetase (ThrRS) was investigated through both in vitro functional experiments and bacterial growth studies. The measured specificity constant (k(cat)/K(M)) for HNV was found to be only 20-30-fold less than that of cognate threonine. The rate of aminoacyl transfer (10.4 s(-1)) was 10-fold higher than the multiple turnover k(cat) value (1 s(-1)), indicating that, as for cognate threonine, amino acid activation is likely to be the rate-limiting step. Like noncognate serine, HNV enhances the ATPase function of the synthetic site, at a rate not increased by nonaminoacylatable (3'-dA76) tRNA. ThrRS also failed to exhibit posttransfer editing activity against HNV. In growing bacteria, the addition of HNV dramatically suppressed growth rates, which indicates either negative phenotypic consequences associated with its inc...Continue Reading

References

Mar 30, 1995·International Journal of Radiation Oncology, Biology, Physics·P MauchH J Deeg
Aug 1, 1995·Toxicology in Vitro : an International Journal Published in Association with BIBRA·G E SchoetersR Van Den Heuvel
Oct 1, 1998·Environmental Toxicology and Pharmacology·L GribaldoE Marafante
Oct 19, 2012·Biochemistry·John J Perona, Andrew Hadd
Oct 8, 2016·Methods : a Companion to Methods in Enzymology·Nevena Cvetesic, Ita Gruic-Sovulj
Oct 29, 2017·Proceedings of the National Academy of Sciences of the United States of America·Jonathan L McMurry, Michelle C Y Chang
Jan 24, 2019·The Journal of Biological Chemistry·Christopher S Francklyn, Patrick Mullen

Citations

Jul 10, 1981·Nucleic Acids Research·H Jakubowski, A R Fersht
Jul 1, 1995·Nature Structural Biology·L MosyakM G Safro
Jul 6, 2000·Nature Structural Biology·R SankaranarayananDino Moras
Aug 2, 2000·Proceedings of the National Academy of Sciences of the United States of America·P J Beuning, K Musier-Forsyth
Aug 31, 2000·Annual Review of Biochemistry·Michael Ibba, D Soll
Sep 24, 2002·The Journal of Biological Chemistry·Leslie A NanglePaul Schimmel
May 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·V PezoValérie de Crécy-Lagard
Apr 23, 2005·The Journal of Biological Chemistry·Nathan T UterJohn J Perona
Mar 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Matthew C T HartmanJack W Szostak
May 13, 2006·Annual Review of Biophysics and Biomolecular Structure·Lei WangPeter G Schultz
Dec 23, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jiqiang LingMichael Ibba
Jul 10, 2007·Journal of Bacteriology·Jamie M BacherPaul Schimmel
Oct 11, 2007·Biochemistry·William F Waas, Paul Schimmel
Aug 30, 2008·The Journal of Biological Chemistry·Yeeting E ChongPaul Schimmel
Nov 11, 2008·Proceedings of the National Academy of Sciences of the United States of America·Anand Minajigi, Christopher S Francklyn
Nov 21, 2008·Proceedings of the National Academy of Sciences of the United States of America·Michal T BonieckiSusan A Martinis
Apr 22, 2009·Annual Review of Microbiology·Jiqiang LingMichael Ibba
May 9, 2009·Proceedings of the National Academy of Sciences of the United States of America·Masahiro NaganumaShigeyuki Yokoyama
Jun 25, 2009·Proceedings of the National Academy of Sciences of the United States of America·Masaaki SokabeIsao Tanaka
Mar 24, 2010·Annual Review of Biochemistry·Chang C Liu, Peter G Schultz
May 26, 2010·The Journal of Biological Chemistry·Morana DulicIta Gruic-Sovulj
May 28, 2010·The Journal of Biological Chemistry·Anand Minajigi, Christopher S Francklyn
Nov 17, 2010·Nature Reviews. Microbiology·Noah M ReynoldsMichael Ibba

Related Concepts

Metabolic Process, Cellular
Activation of Amino Acids
Alkalescens-Dispar Group
Adenosine Triphosphatases
Amino Acids, I.V. solution additive
MT-TA gene
Triplet Codon-amino Acid Adaptor Activity
Transfer RNA
Protein Biosynthesis
Threonine-tRNA Ligase

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