1887

Microbiology

Volume 155, Issue 6

Homocysteine editing and growth inhibition in Free

Abstract

In homocysteine (Hcy) is metabolically converted to the thioester Hcy-thiolactone in ATP-consuming reactions catalysed by methionyl-, isoleucyl- and leucyl-tRNA synthetases. Here we show that growth inhibition caused by supplementation of cultures with Hcy is accompanied by greatly increased accumulation of Hcy-thiolactone. Energy dissipation for Hcy editing increases 100-fold in the presence of exogenous Hcy and reaches one mole of ATP unproductively dissipated for Hcy-thiolactone synthesis per each mole of ATP that is consumed for methionine activation. Inhibiting Hcy-thiolactone synthesis with isoleucine, leucine or methionine accelerates bacterial growth in Hcy-supplemented cultures. Growth rates in Hcy-inhibited cultures are inversely related to the accumulation of Hcy-thiolactone. We also show that the levels of protein -linked Hcy modestly increase in cells in Hcy-supplemented cultures. The results suggest that Hcy editing restrains bacterial growth.

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  • Published Online:
Keyword(s): DTT, dithiothreitol , Hcy, homocysteine , IleRS, isoleucyl-tRNA synthetase , LeuRS, leucyl-tRNA synthetase and MetRS, methionyl-tRNA synthetase
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2009-06-01
2019-10-14

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References

  1. Chwatko, G. & Jakubowski, H. ( 2005; ). The determination of homocysteine-thiolactone in human plasma. Anal Biochem 337, 271–277.[CrossRef]
    [Google Scholar]
  2. Chwatko, G., Boers, G. H., Strauss, K. A., Shih, D. M. & Jakubowski, H. ( 2007; ). Mutations in methylenetetrahydrofolate reductase or cystathionine beta-synthase gene, or a high-methionine diet, increase homocysteine thiolactone levels in humans and mice. FASEB J 21, 1707–1713.[CrossRef]
    [Google Scholar]
  3. Gao, W., Goldman, E. & Jakubowski, H. ( 1994; ). Role of carboxy-terminal region in proofreading function of methionyl-tRNA synthetase in Escherichia coli. Biochemistry 33, 11528–11535.[CrossRef]
    [Google Scholar]
  4. Ingraham, J. L., Maaløe, O. & Neidhardt, F. C. ( 1983; ). Growth of the Bacterial Cell. Sunderland, MA: Sinauer Associates.
  5. Jakubowski, H. ( 1990; ). Proofreading in vivo: editing of homocysteine by methionyl-tRNA synthetase in Escherichia coli. Proc Natl Acad Sci U S A 87, 4504–4508.[CrossRef]
    [Google Scholar]
  6. Jakubowski, H. ( 1991; ). Proofreading in vivo: editing of homocysteine by methionyl-tRNA synthetase in the yeast Saccharomyces cerevisiae. EMBO J 10, 593–598.
     [Google Scholar]
  7. Jakubowski, H. ( 1993; ). Energy cost of proofreading in vivo: the charging of methionine tRNAs in Escherichia coli. FASEB J 7, 168–172.
     [Google Scholar]
  8. Jakubowski, H. ( 1995; ). Proofreading in vivo. Editing of homocysteine by aminoacyl-tRNA synthetases in Escherichia coli. J Biol Chem 270, 17672–17673.
     [Google Scholar]
  9. Jakubowski, H. ( 1997a; ). Metabolism of homocysteine thiolactone in human cell cultures. Possible mechanism for pathological consequences of elevated homocysteine levels. J Biol Chem 272, 1935–1942.
     [Google Scholar]
  10. Jakubowski, H. ( 1997b; ). Aminoacyl thioester chemistry of class II aminoacyl-tRNA synthetases. Biochemistry 36, 11077–11085.[CrossRef]
    [Google Scholar]
  11. Jakubowski, H. ( 1999a; ). Misacylation of tRNALys with noncognate amino acids by lysyl-tRNA synthetase. Biochemistry 38, 8088–8093.[CrossRef]
    [Google Scholar]
  12. Jakubowski, H. ( 1999b; ). Protein homocysteinylation: possible mechanism underlying pathological consequences of elevated homocysteine levels. FASEB J 13, 2277–2283.
     [Google Scholar]
  13. Jakubowski, H. ( 2000; ). Translational incorporation of S-nitrosohomocysteine into protein. J Biol Chem 275, 21813–21816.[CrossRef]
    [Google Scholar]
  14. Jakubowski, H. ( 2001a; ). Protein N-homocysteinylation: implications for atherosclerosis. Biomed Pharmacother 55, 443–447.[CrossRef]
    [Google Scholar]
  15. Jakubowski, H. ( 2001b; ). Translational accuracy of aminoacyl-tRNA synthetases: implications for atherosclerosis. J Nutr 131, 2983S–2987S.
     [Google Scholar]
  16. Jakubowski, H. ( 2002a; ). The determination of homocysteine-thiolactone in biological samples. Anal Biochem 308, 112–119.[CrossRef]
    [Google Scholar]
  17. Jakubowski, H. ( 2002b; ). Homocysteine is a protein amino acid in humans. Implications for homocysteine-linked disease. J Biol Chem 277, 30425–30428.[CrossRef]
    [Google Scholar]
  18. Jakubowski, H. ( 2004; ). Molecular basis of homocysteine toxicity in humans. Cell Mol Life Sci 61, 470–487.[CrossRef]
    [Google Scholar]
  19. Jakubowski, H. ( 2005a; ). tRNA synthetase editing of amino acids. In Encyclopedia of Life Sciences http://mrw.interscience.wiley.com/emrw/9780470015902/els/article/a0000532/current/abstract doi:10.1038/npg.els.0003933.
  20. Jakubowski, H. ( 2005b; ). Accuracy of aminoacyl-tRNA synthetases: proofreading of amino acids. In The Aminoacyl-tRNA Synthetases, pp. 384–396. Edited by M. Ibba, C. Francklyn & S. Cusack. Georgetown, TX: Landes Bioscience/Eurekah.com.
  21. Jakubowski, H. ( 2006a; ). Pathophysiological consequences of homocysteine excess. J Nutr 136, 1741S–1749S.
     [Google Scholar]
  22. Jakubowski, H. ( 2006b; ). Mechanism of the condensation of homocysteine thiolactone with aldehydes. Chemistry 12, 8039–8043.[CrossRef]
    [Google Scholar]
  23. Jakubowski, H. ( 2007a; ). The molecular basis of homocysteine thiolactone-mediated vascular disease. Clin Chem Lab Med 45, 1704–1716.
     [Google Scholar]
  24. Jakubowski, H. ( 2007b; ). Facile syntheses of [35S]homocysteine-thiolactone, [35S]homocystine, [35S]homocysteine, and [S-nitroso-35S]homocysteine. Anal Biochem 370, 124–126.[CrossRef]
    [Google Scholar]
  25. Jakubowski, H. ( 2008; ). New method for the determination of protein N-linked homocysteine. Anal Biochem 380, 257–261.[CrossRef]
    [Google Scholar]
  26. Jakubowski, H. & Fersht, A. R. ( 1981; ). Alternative pathways for editing non-cognate amino acids by aminoacyl-tRNA synthetases. Nucleic Acids Res 9, 3105–3117.[CrossRef]
    [Google Scholar]
  27. Jakubowski, H. & Goldman, E. ( 1992; ). Editing of errors in selection of amino acids for protein synthesis. Microbiol Rev 56, 412–429.
     [Google Scholar]
  28. Karkhanis, V. A., Mascarenhas, A. P. & Martinis, S. A. ( 2007; ). Amino acid toxicities of Escherichia coli that are prevented by leucyl-tRNA synthetase amino acid editing. J Bacteriol 189, 8765–8768.[CrossRef]
    [Google Scholar]
  29. Kumar, A., John, L., Alam, M. M., Gupta, A., Sharma, G., Pillai, B. & Sengupta, S. ( 2006; ). Homocysteine- and cysteine-mediated growth defect is not associated with induction of oxidative stress response genes in yeast. Biochem J 396, 61–69.[CrossRef]
    [Google Scholar]
  30. Mulvey, R. S. & Fersht, A. R. ( 1977; ). Editing mechanisms in aminoacylation of tRNA: ATP consumption and the binding of aminoacyl-tRNA by elongation factor Tu. Biochemistry 16, 4731–4737.[CrossRef]
    [Google Scholar]
  31. Roe, A. J., O'Byrne, C., McLaggan, D. & Booth, I. R. ( 2002; ). Inhibition of Escherichia coli growth by acetic acid: a problem with methionine biosynthesis and homocysteine toxicity. Microbiology 148, 2215–2222.
     [Google Scholar]
  32. Teusink, B., van Enckevort, F. H., Francke, C., Wiersma, A., Wegkamp, A., Smid, E. J. & Siezen, R. J. ( 2005; ). In silico reconstruction of the metabolic pathways of Lactobacillus plantarum: comparing predictions of nutrient requirements with those from growth experiments. Appl Environ Microbiol 71, 7253–7262.[CrossRef]
    [Google Scholar]
  33. Tuite, N. L., Fraser, K. R. & O'Byrne, C. P. ( 2005; ). Homocysteine toxicity in Escherichia coli is caused by a perturbation of branched-chain amino acid biosynthesis. J Bacteriol 187, 4362–4371.[CrossRef]
    [Google Scholar]
  34. Zimny, J., Sikora, M., Guranowski, A. & Jakubowski, H. ( 2006; ). Protective mechanisms against homocysteine toxicity: the role of bleomycin hydrolase. J Biol Chem 281, 22485–22492.[CrossRef]
    [Google Scholar]
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Homocysteine editing and growth inhibition in Escherichia coli
Microbiology 155, 1858 (2009); https://doi.org/10.1099/mic.0.026609-0
/content/journal/micro/10.1099/mic.0.026609-0
/content/journal/micro/10.1099/mic.0.026609-0
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