Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1994 Apr 11;22(7):1155–1160. doi: 10.1093/nar/22.7.1155

Editing function of Escherichia coli cysteinyl-tRNA synthetase: cyclization of cysteine to cysteine thiolactone.

H Jakubowski 1
PMCID: PMC523636  PMID: 8165127

Abstract

A cyclic sulfur compound, identified as cysteine thiolactone by several chemical and enzymatic tests, is formed from cysteine during in vitro tRNA(Cys) aminoacylation catalyzed by Escherichia coli cysteinyl-tRNA synthetase. The mechanism of cysteine thiolactone formation involves enzymatic deacylation of Cys-tRNA(Cys) (k = 0.017 s-1) in which nucleophilic sulfur of the side chain of cysteine in Cys-tRNA(Cys) attacks its carboxyl carbon to yield cysteine thiolactone. Nonenzymatic deacylation of Cys-tRNA(Cys) (k = 0.0006 s-1) yields cysteine, as expected. Inhibition of enzymatic deacylation of Cys-tRNA(Cys) by cysteine and Cys-AMP, but not by ATP, indicates that both synthesis of Cys-tRNA(Cys) and cyclization of cysteine to the thiolactone occur in a single active site of the enzyme. The cyclization of cysteine is mechanistically similar to the editing reactions of methionyl-tRNA synthetase. However, in contrast to methionyl-tRNA synthetase which needs the editing function to reject misactivated homocysteine, cysteinyl-tRNA synthetase is highly selective and is not faced with a problem in rejecting noncognate amino acids. Despite this, the present day cysteinyl-tRNA synthetase, like methionyl-tRNA synthetase, still retains an editing activity toward the cognate product, the charged tRNA. This function may be a remnant of a chemistry used by an ancestral cysteinyl-tRNA synthetase.

Full text

PDF
1155

Images in this article

1156Image
on p.1156
1157Image
on p.1157
1157Image
on p.1157

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avalos J., Corrochano L. M., Brenner S. Cysteinyl-tRNA synthetase is a direct descendant of the first aminoacyl-tRNA synthetase. FEBS Lett. 1991 Jul 29;286(1-2):176–180. doi: 10.1016/0014-5793(91)80968-9. [DOI] [PubMed] [Google Scholar]
  2. Eriani G., Dirheimer G., Gangloff J. Cysteinyl-tRNA synthetase: determination of the last E. coli aminoacyl-tRNA synthetase primary structure. Nucleic Acids Res. 1991 Jan 25;19(2):265–269. doi: 10.1093/nar/19.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fersht A. R., Dingwall C. An editing mechanism for the methionyl-tRNA synthetase in the selection of amino acids in protein synthesis. Biochemistry. 1979 Apr 3;18(7):1250–1256. doi: 10.1021/bi00574a021. [DOI] [PubMed] [Google Scholar]
  4. Fersht A. R., Dingwall C. Cysteinyl-tRNA synthetase from Escherichia coli does not need an editing mechanism to reject serine and alanine. High binding energy of small groups in specific molecular interactions. Biochemistry. 1979 Apr 3;18(7):1245–1249. doi: 10.1021/bi00574a020. [DOI] [PubMed] [Google Scholar]
  5. Fersht A. R., Dingwall C. Establishing the misacylation/deacylation of the tRNA pathway for the editing mechanism of prokaryotic and eukaryotic valyl-tRNA synthetases. Biochemistry. 1979 Apr 3;18(7):1238–1245. doi: 10.1021/bi00574a019. [DOI] [PubMed] [Google Scholar]
  6. Hou Y. M., Shiba K., Mottes C., Schimmel P. Sequence determination and modeling of structural motifs for the smallest monomeric aminoacyl-tRNA synthetase. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):976–980. doi: 10.1073/pnas.88.3.976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jakubowski H. Z., Pastuzyn A., Loftfield R. B. The determination of aminoacyl adenylate by thin-layer chromatography. Anal Biochem. 1977 Sep;82(1):29–37. doi: 10.1016/0003-2697(77)90130-0. [DOI] [PubMed] [Google Scholar]
  8. Jakubowski H. Energy cost of proofreading in vivo: the charging of methionine tRNAs in Escherichia coli. FASEB J. 1993 Jan;7(1):168–172. doi: 10.1096/fasebj.7.1.8422964. [DOI] [PubMed] [Google Scholar]
  9. Jakubowski H., Fersht A. R. Alternative pathways for editing non-cognate amino acids by aminoacyl-tRNA synthetases. Nucleic Acids Res. 1981 Jul 10;9(13):3105–3117. doi: 10.1093/nar/9.13.3105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jakubowski H., Goldman E. Editing of errors in selection of amino acids for protein synthesis. Microbiol Rev. 1992 Sep;56(3):412–429. doi: 10.1128/mr.56.3.412-429.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jakubowski H., Goldman E. Synthesis of homocysteine thiolactone by methionyl-tRNA synthetase in cultured mammalian cells. FEBS Lett. 1993 Feb 15;317(3):237–240. doi: 10.1016/0014-5793(93)81283-6. [DOI] [PubMed] [Google Scholar]
  12. Jakubowski H. Proofreading and the evolution of a methyl donor function. Cyclization of methionine to S-methyl homocysteine thiolactone by Escherichia coli methionyl-tRNA synthetase. J Biol Chem. 1993 Mar 25;268(9):6549–6553. [PubMed] [Google Scholar]
  13. Jakubowski H. Proofreading in vivo: editing of homocysteine by methionyl-tRNA synthetase in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4504–4508. doi: 10.1073/pnas.87.12.4504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jakubowski H. Proofreading in vivo: editing of homocysteine by methionyl-tRNA synthetase in the yeast Saccharomyces cerevisiae. EMBO J. 1991 Mar;10(3):593–598. doi: 10.1002/j.1460-2075.1991.tb07986.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kim H. Y., Ghosh G., Schulman L. H., Brunie S., Jakubowski H. The relationship between synthetic and editing functions of the active site of an aminoacyl-tRNA synthetase. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11553–11557. doi: 10.1073/pnas.90.24.11553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Old J. M., Jones D. S. The aminoacylation of transfer ribonucleic acid. Inhibitory effects of some amino acid analogues with altered side chains. Biochem J. 1976 Dec 1;159(3):503–511. doi: 10.1042/bj1590503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schreier A. A., Schimmel P. R. Transfer ribonucleic acid synthetase catalyzed deacylation of aminoacyl transfer ribonucleic acid in the absence of adenosine monophosphate and pyrophosphate. Biochemistry. 1972 Apr 25;11(9):1582–1589. doi: 10.1021/bi00759a006. [DOI] [PubMed] [Google Scholar]
  18. Smith L. T., Cohn M. Role of the beta-phosphate-gamma-phosphate interchange reaction of adenosine triphosphate in amino acid discrimination by valyl- and methionyl-tRNA synthetases from Escherichia coli. Biochemistry. 1981 Jan 20;20(2):385–391. doi: 10.1021/bi00505a025. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES