PMID: 2113914Jul 1, 1990Paper

Overproduction of tyrosyl-tRNA synthetase is toxic to Escherichia coli: a genetic analysis

Journal of Bacteriology
Hugues BedouelleM Hermann

Abstract

The tyrS genes from Escherichia coli and Bacillus stearothermophilus were toxic to E. coli when they were carried by plasmids with very high copy numbers (pEMBL8 and pEMBL9). We quantified this effect by comparing the efficiencies of plating of E. coli derivatives harboring recombinant plasmids in various experimental conditions. The toxicity was apparent at both 30 and 37 degrees C. It increased with the growth temperature, the strength of the tyrS promoter, and the copy number of the plasmidic vector. Two- to threefold enhancement of tyrS expression raised the toxicity 300-fold. Point mutations in tyrS that prevent interaction between its product, tyrosyl-tRNA synthetase, and tRNA(Tyr) but do not alter the rate of formation of tyrosyl-adenylate abolished the toxicity. Thus, the toxic effect was due to high cellular levels of synthetase activity. At 30 degrees C, the cellular concentration of tyrosyl-tRNA synthetase reached 55% of that of soluble proteins and led to decreased beta-galactosidase stability. We discuss possible causes of this toxic effect and describe its applications to the study of the recognition and interaction between the synthetase and tRNA(Tyr).

References

Feb 1, 1990·Research in Microbiology·M Hermann, H Bedouelle
Feb 20, 1989·Journal of Molecular Biology·E Labouze, H Bedouelle
Jan 1, 1987·Methods in Enzymology·L Dente, R Cortese
May 31, 1972·Nature: New Biology·B Hall, J Gallant
Jan 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·H A de BoerM Vasser
May 2, 1983·European Journal of Biochemistry·G WinterD G Barker

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Citations

Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·P Schimmel, L Ribas de Pouplana
May 29, 1999·The Journal of Biological Chemistry·C C Wang, P Schimmel
Mar 25, 2009·FEMS Microbiology Letters·Sonja Lesjak, Ivana Weygand-Durasevic
Jan 15, 2015·Proceedings of the National Academy of Sciences of the United States of America·Anna V SherwoodTina M Henkin
May 18, 2007·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Robert A HoltDuane E Smailus
Jan 1, 1993·Biochimie·H BedouelleR Nageotte
Dec 1, 1993·Microbiological Reviews·M Riley

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