Non-orthologous Replacement of Lysyl-tRNA Synthetase Prevents Addition of Lysine Analogs to the Genetic Code

Authors

Brian C. Jester, Trinity College
Jeffrey D. Levengood, The Ohio State University
Hervé Roy, The Ohio State University
Michael Ibba, Chapman UniversityFollow
Kevin M. Devine, Trinity College

Document Type

Article

Publication Date

11-25-2003

Abstract

Insertion of lysine during protein synthesis depends on the enzyme lysyl-tRNA synthetase (LysRS), which exists in two unrelated forms, LysRS1 and LysRS2. LysRS1 has been found in most archaea and some bacteria, and LysRS2 has been found in eukarya, most bacteria, and a few archaea, but the two proteins are almost never found together in a single organism. Comparison of structures of LysRS1 and LysRS2 complexed with lysine suggested significant differences in their potential to bind lysine analogues with backbone replacements. One such naturally occurring compound, the metabolic intermediate S-(2-aminoethyl)-l-cysteine, is a bactericidal agent incorporated during protein synthesis via LysRS2. In vitro tests showed that S-(2-aminoethyl)-l-cysteine is a poor substrate for LysRS1, and that it inhibits LysRS1 200-fold less effectively than it inhibits LysRS2. In vivo inhibition by S-(2-aminoethyl)-l-cysteine was investigated by replacing the endogenous LysRS2 of Bacillus subtilis with LysRS1 from the Lyme disease pathogen Borrelia burgdorferi. B. subtilis strains producing LysRS1 alone were relatively insensitive to growth inhibition by S-(2-aminoethyl)-l-cysteine, whereas a WT strain or merodiploid strains producing both LysRS1 and LysRS2 showed significant growth inhibition under the same conditions. These growth effects arising from differences in amino acid recognition could contribute to the distribution of LysRS1 and LysRS2 in different organisms. More broadly, these data demonstrate how diversity of the aminoacyl-tRNA synthetases prevents infiltration of the genetic code by noncanonical amino acids, thereby providing a natural reservoir of potential antibiotic resistance.

Comments

This article was originally published in Proceedings of the National Academy of Sciences of the United States of America, volume 100, in 2003. https://doi.org/10.1073/pnas.2036253100

Recommended Citation

Jester, B., Levengood, J., Roy, H., Ibba, M. and Devine, K. (2003) Non-orthologous replacement of lysyl-tRNA synthetase prevents addition of lysine analogs to the genetic code. Proc. Natl. Acad. Sci. USA 100, 14351-14356. https://doi.org/10.1073/pnas.2036253100

Copyright

The National Academy of Sciences