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Volume 152, Issue 8

Global effects of homocysteine on transcription in : induction of the gene for the major cold-shock protein, CspA Free

Abstract

Homocysteine (Hcy) is a thiol-containing amino acid that is considered to be medically important because it is linked to the development of several life-threatening diseases in humans, including cardiovascular disease and stroke. It inhibits the growth of when supplied in the growth medium. Growth inhibition is believed to arise as a result of partial starvation for isoleucine, which occurs because Hcy perturbs the biosynthesis of this amino acid. This study attempted to further elucidate the inhibitory mode of action of Hcy by examining the impact of exogenously supplied Hcy on the transcriptome. Using gene macroarrays the transcript levels corresponding to 68 genes were found to be reproducibly altered in the presence of 0.5 mM Hcy. Of these genes, the biggest functional groups affected were those involved in translation (25 genes) and in amino acid metabolism (19 genes). Genes involved in protection against oxidative stress were repressed in Hcy-treated cells and this correlated with a decrease in catalase activity. The gene showing the strongest induction by Hcy was , which encodes the major cold-shock protein CspA. RT-PCR and reporter fusion experiments confirmed that was induced by Hcy. Induction of by Hcy was not caused by nutritional upshift, a stimulus known to induce CspA expression, nor was it dependent on the presence of a functional CspA protein. The induction of by Hcy was suppressed when isoleucine was included in the growth medium. These data suggest that the induction of CspA expression in the presence of Hcy occurs because of a limitation for isoleucine. The possibility that Hcy-induced expression is triggered by translational stalling that occurs when the cells are limited for isoleucine is discussed.

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Keyword(s): BCAA, branched-chain amino acid , Hcy, homocysteine and THF, tetrahydrofolate
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2006-08-01
2024-04-18
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References

  1. Adams M. D, Wagner L. M, Graddis T. J, Landick R, Antonucci T. K, Gibson A. L, Oxender D. L. 1990; Nucleotide sequence and genetic characterisation reveal six essential genes for the LIV-1 and LS transport systems of Escherichia coli . J Biol Chem 265:11436–11443
     [Google Scholar]
  2. Anderson J. L, Muhlestein J. B, Horne B. D, Carlquist J. F, Bair T. L, Madsen T. E, Pearson R. R. 2000; Plasma homocysteine predicts mortality independently of traditional risk factors and C-reactive protein in patients with angiographically defined coronary artery disease. Circulation 102:1227–1232 [CrossRef]
     [Google Scholar]
  3. Bae W, Jones P. G, Inouye M. 1997; CspA, the major cold shock protein of Escherichia coli , negatively regulates its own gene expression. J Bacteriol 179:7081–7088
     [Google Scholar]
  4. Beers R. F., Jr, Sizer I. W. 1952; A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–140
     [Google Scholar]
  5. Berglin E. H, Edlund M. B, Nyberg G. K, Carlsson J. 1982; Potentiation by l-cysteine of the bactericidal effect of hydrogen peroxide in Escherichia coli . J Bacteriol 152:81–88
     [Google Scholar]
  6. Bhagwat A. A, Phadke R. P, Wheeler D, Kalantre S, Gudipati M, Bhagwat M. 2003; Computational methods and evaluation of RNA stabilization reagents for genome-wide expression studies. J Microbiol Methods 55:399–409 [CrossRef]
     [Google Scholar]
  7. Bianchi A. A, Baneyx F. 1999; Stress responses as a tool to detect and characterize the mode of action of antibacterial agents. Appl Environ Microbiol 65:5023–5027
     [Google Scholar]
  8. Brandi A, Spurio R, Gualerzi C. O, Pon C. L. 1999; Massive presence of the Escherichia coli ‘major cold-shock protein’ CspA under non-stress conditions. EMBO J 18:1653–1659 [CrossRef]
     [Google Scholar]
  9. Byerly K. A, Urbanowski M. L, Stauffer G. V. 1990; Escherichia coli metR mutants that produce a MetR activator protein with an altered homocysteine response. J Bacteriol 172:2839–2843
     [Google Scholar]
  10. Bystrom A. S, Bjork G. R. 1982; Chromosomal location and cloning of the gene (trmD) responsible for the synthesis of tRNA (m1G) methyltransferase in Escherichia coli K-12. Mol Gen Genet 188:440–446 [CrossRef]
     [Google Scholar]
  11. Cai X. Y, Maxon M, Redfield B, Glass R. E, Brot N, Weissbach H. 1989; Methionine synthesis in Escherichia coli : effect of the MetR protein on metE and metH expression. Proc Natl Acad Sci U S A 86:4407–4411 [CrossRef]
     [Google Scholar]
  12. Casas J. P, Bautista L. E, Smeeth L, Sharma P, Hingorani A. D. 2005; Homocysteine and stroke: evidence on a causal link from mendelian randomisation. Lancet 365:224–232 [CrossRef]
     [Google Scholar]
  13. De Groote M. A, Testerman T, Xu Y, Stauffer G, Fang F. C. 1996; Homocysteine antagonism of nitric oxide-related cytostasis in Salmonella typhimurium . Science 272:414–417 [CrossRef]
     [Google Scholar]
  14. Goldstein J, Lehnhardt S, Inouye M. 1990; Major cold shock protein of Escherichia coli . Proc Natl Acad Sci U S A 87:283–287 [CrossRef]
     [Google Scholar]
  15. Graumann P, Wendrich T. M, Weber M. H, Schroder K, Marahiel M. A. 1997; A family of cold shock proteins in Bacillus subtilis is essential for cellular growth and for efficient protein synthesis at optimal and low temperatures. Mol Microbiol 25:741–756 [CrossRef]
     [Google Scholar]
  16. Greene R. C. others 1996; Biosynthesis of methioinine. In Escherichia coli and Salmonella: Cellular and Molecular Biology pp  514–527 Edited by Neidhardt F. C. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  17. Harris C. L. 1981; Cysteine and growth inhibition of Escherichia coli : threonine deaminase as the target enzyme. J Bacteriol 145:1031–1035
     [Google Scholar]
  18. Huang L. E, Zhang H, Bae S. W, Liu A. Y. 1994; Thiol reducing reagents inhibit the heat shock response. Involvement of a redox mechanism in the heat shock signal transduction pathway. J Biol Chem 269:30718–30725
     [Google Scholar]
  19. Jakubowski H. 1999; Protein homocysteinylation: possible mechanism underlying pathological consequences of elevated homocysteine levels. FASEB J 13:2277–2283
     [Google Scholar]
  20. Jakubowski H. 2000; Translational incorporation of S -nitrosohomocysteine into protein. J Biol Chem 275:21813–21816 [CrossRef]
     [Google Scholar]
  21. Jakubowski H. 2002; The determination of homocysteine-thiolactone in biological samples. Anal Biochem 308:112–119 [CrossRef]
     [Google Scholar]
  22. Jakubowski H. 2004; Molecular basis of homocysteine toxicity in humans. Cell Mol Life Sci 61:470–487 [CrossRef]
     [Google Scholar]
  23. Jiang W, Jones P, Inouye M. 1993; Chloramphenicol induces the transcription of the major cold shock gene of Escherichia coli , cspA . J Bacteriol 175:5824–5828
     [Google Scholar]
  24. Kredich N. M. others 1996; Biosynthesis of cysteine. In Escherichia coli and Salmonella: Cellular and Molecular Biology pp  514–527 Edited by Neidhardt F. C. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  25. Loscalzo J. 1996; The oxidant stress of hyperhomocyst(e)inemia. J Clin Invest 98:5–7 [CrossRef]
     [Google Scholar]
  26. Lynch S. M, Frei B. 1997; Physiological thiol compounds exert pro- and anti-oxidant effects, respectively, on iron- and copper-dependent oxidation of human low-density lipoprotein. Biochim Biophys Acta 1345215–221 [CrossRef]
     [Google Scholar]
  27. Maki Y, Yoshida H, Wada A. 2000; Two proteins, YfiA and YhbH, associated with resting ribosomes in stationary phase Escherichia coli . Genes Cells 5:965–974 [CrossRef]
     [Google Scholar]
  28. Merlin C, Gardiner G, Durand S, Masters M. 2002; The Escherichia coli metD locus encodes an ABC transporter which includes Abc (MetN),YaeE (MetI), and YaeC (MetQ). J Bacteriol 184:5513–5517 [CrossRef]
     [Google Scholar]
  29. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  30. Miller J. H. 1992 A Short Course in Bacterial Genetics: a Laboratory Manual and Handbook for Escherichia coli and Related Bacteria Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  31. Mitta M, Fang L, Inouye M. 1997; Deletion analysis of cspA of Escherichia coli : requirement of the AT-rich UP element for cspA transcription and the downstream box in the coding region for its cold shock induction. Mol Microbiol 26:321–335 [CrossRef]
     [Google Scholar]
  32. Outinen P. A, Sood S. K, Liaw P. C. 7 other authors 1998; Characterization of the stress-inducing effects of homocysteine. Biochem J 332:213–221
     [Google Scholar]
  33. Park S, Imlay J. A. 2003; High levels of intracellular cysteine promote oxidative DNA damage by driving the Fenton reaction. J Bacteriol 185:1942–1950 [CrossRef]
     [Google Scholar]
  34. Phadtare S, Yamanaka K, Inouye M. 2000; The cold shock response. In Bacterial Stress Responses pp  33–45 Edited by Storz G., Hengge-Aronis R. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  35. 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]
  36. Seshadri S, Beiser A, Selhub J, Jacques P. F, Rosenberg I. H, D'Agostino R. B, Wilson P. W, Wolf P. A. 2002; Plasma homocysteine as a risk factor for dementia and Alzheimer's disease. N Engl J Med 346:476–483 [CrossRef]
     [Google Scholar]
  37. Stamler J. S, Osborne J. A, Jaraki O, Rabbani L. E, Mullins M, Singel D, Loscalzo J. 1993; Adverse vascular effects of homocysteine are modulated by endothelium-derived relaxing factor and related oxides of nitrogen. J Clin Invest 91:308–318 [CrossRef]
     [Google Scholar]
  38. Starkebaum G, Harlan J. M. 1986; Endothelial cell injury due to copper-catalyzed hydrogen peroxide generation from homocysteine. J Clin Invest 77:1370–1376 [CrossRef]
     [Google Scholar]
  39. 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]
  40. Umbarger H. E. others 1996; Biosynthesis of the branched-chain amino acids. In Escherichi coli and Salmonella: Cellular and Molecular Biology pp  442–457 Edited by Neidhardt F. C. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  41. Upchurch G. R., Jr, Welch G. N, Fabian A. J, Freedman J. E, Johnson J. L, Loscalzo J, Keaney J. F., Jr. 1997; Homocyst(e)ine decreases bioavailable nitric oxide by a mechanism involving glutathione peroxidase. J Biol Chem 272:17012–17017 [CrossRef]
     [Google Scholar]
  42. Urbanowski M. L, Stauffer G. V. 1989; Role of homocysteine in metR -mediated activation of the metE and metH genes in Salmonella typhimurium and Escherichia coli . J Bacteriol 171:3277–3281
     [Google Scholar]
  43. VanBogelen R. A, Neidhardt F. C. 1990; Ribosomes as sensors of heat and cold shock in Escherichia coli . Proc Natl Acad Sci U S A 87:5589–5593 [CrossRef]
     [Google Scholar]
  44. van der Put N. M, van Straaten H. W, Trijbels F. J, Blom H. J. 2001; Folate, homocysteine and neural tube defects: an overview. Exp Biol Med (Maywood) 226:243–270
     [Google Scholar]
  45. Vollset S. E, Refsum H, Irgens L. M, Emblem B. M, Tverdal A, Gjessing H. K, Monsen A. L, Ueland P. M. 2000; Plasma total homocysteine, pregnancy complications, and adverse pregnancy outcomes: the Hordaland Homocysteine study. Am J Clin Nutr 71:962–968
     [Google Scholar]
  46. Winzer K, Hardie K. R, Burgess N. 8 other authors 2002; LuxS: its role in central metabolism and the in vitro synthesis of 4-hydroxy-5-methyl-3(2H)-furanone. Microbiology 148:909–922
     [Google Scholar]
  47. Wu W. F, Urbanowski M. L, Stauffer G. V. 1995; Characterization of a second MetR-binding site in the metE metR regulatory region of Salmonella typhimurium . J Bacteriol 177:1834–1839
     [Google Scholar]
  48. Yamanaka K, Inouye M. 2001; Induction of CspA, an E. coli major cold-shock protein, upon nutritional upshift at 37 °C. Genes Cells 6:279–290 [CrossRef]
     [Google Scholar]
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Global effects of homocysteine on transcription in Escherichia coli: induction of the gene for the major cold-shock protein, CspA
Microbiology 152, 2221 (2006); https://doi.org/10.1099/mic.0.28804-0
/content/journal/micro/10.1099/mic.0.28804-0
/content/journal/micro/10.1099/mic.0.28804-0
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