1887

Abstract

Alginate production and degree of polymerization were affected when the highly mucoid 8821M was grown with growth-inhibitory concentrations of Cu (supplied as CuCl; 1-5 m). The inhibition of alginate biosynthesis was consistent with the decreased activity in Cu-stressed cells of phosphomannose isomerase/GDP-mannose pyrophosphorylase (encoded by ), phosphomannomutase (encoded by ) and GDP-mannose dehydrogenase (encoded by ). However, in cells grown with concentrations of CuCl below 2 m the steady-state mRNA levels from and from the regulatory gene increased moderately. This observation is consistent with the suggested linkage between the control of alginate gene expression and the global regulation involved in the oxidative stress response. At highly inhibitory concentrations the levels of the four alginate gene transcripts decreased from maximal values. The bell-shaped curves, representing the effect of Cu concentration on mRNA levels from the four alginate genes, exhibited similar patterns but did not concur. The decrease of the specific activity of enzymes necessary for GDP-mannuronic acid synthesis in Cu-grown cells was correlated with changes in gene expression, with the inhibitory effect of Cu on enzyme activities and with Cu-induced oxidative inactivation of enzymes, especially the particularly sensitive phosphomannose isomerase activity.

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1997-02-01
2021-08-05
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References

  1. Alves M.J., Martins L.O., Sá-Correia I. 1991; Temperature profiles and alginate synthesis in mucoid and non-mucoid variants of Pseudomonas aeruginosa. . Lett Appl Microbiol 12:244–248
    [Google Scholar]
  2. Appanna V.D., Preston C.M. 1987; Manganese elicits the synthesis of a novel exopolysaccharide in an arctic Rhizobium. . FEBS Lett 140:79–82
    [Google Scholar]
  3. Berry A., DeVault J.D., Chakrabarty A.M. 1989; High osmolarity is a signal for enhanced algD transcription in mucoid and nonmucoid Pseudomonas aeruginosa strains.. J Bacteriol 171:2312–2317
    [Google Scholar]
  4. Bradford M.M. 1976; A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
    [Google Scholar]
  5. Chitnis C.E., Ohman D.E. 1993; Genetic analysis of the alginate biosynthetic gene cluster of Pseudomonas aeruginosa shows evidence of an operonic structure.. Mol Microbiol 8:583–590
    [Google Scholar]
  6. Deretic V., Konyescni W.M. 1989; Control of mucoidy in Pseudomonas aeruginosa: transcriptional regulation of algR and identification of the second regulatory gene, algQ. . J Bacteriol 171:3680–3688
    [Google Scholar]
  7. Deretic V., Gill J.F., Chakrabarty A.M. 1987; Gene algD, coding for GDP-mannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa. . J Bacteriol 169:351–358
    [Google Scholar]
  8. Deretic V., Dikshit R., Konyescni W.M., Chakrabarty A.M., Misra T.K. 1989; The algR gene, which regulates mucoidy in Pseudomonas aeruginosa, belongs to a class of environmentally responsive genes.. J Bacteriol 171:1278–1283
    [Google Scholar]
  9. Deretic V., Leveau J.H.J., Mohr C.D., Hibler N.S. 1992; In vitro phosphorylation of AlgR, a regulator of mucoidy in Pseudomonas aeruginosa, by a histidine protein kinase and effects of small phospho-donor molecules.. Mol Microbiol 6:2761–2767
    [Google Scholar]
  10. Deretic V., Schurr M.J., Boucher J.C., Martin D.W. 1994; Conversion of Pseudomonas aeruginosa to mucoidy in cystic fibrosis: environmental stress and regulation of bacterial virulence by alternative sigma factors.. J Bacteriol 176:2773–2780
    [Google Scholar]
  11. DeVries C.A., Ohman D.E. 1994; Mucoid-to-nonmucoid conversion in alginate-producing Pseudomonas aeruginosa often results from spontaneous mutations in algT, encoding a putative alternate sigma factor, and shows evidence for autoregulation.. J Bacteriol 176:6677–6687
    [Google Scholar]
  12. Fujiwara S., Zielinski N.A., Chakrabarty A.M. 1993; Enhancer-like activity of AlgRl-binding site in alginate gene activation: positional, orientational, and sequence specificity.. J Bacteriol 175:5452–5459
    [Google Scholar]
  13. Halliwell B., Gutteridge J.M. 1989 Free Radicals in Biology and Medicine, 2nd edn.. Oxford: Oxford University Press;
    [Google Scholar]
  14. Higgins C.F., Causton H.C., Dance G.S.C., Mudd E.A. 1993; The role of the 3´ end in mRNA stability and decay.. In Control of Messenger RNA Stability pp. 13–30 Edited by Belasco J., Brawerman G. San Diego: Academic Press;
    [Google Scholar]
  15. Kidambi S.P., Sundin G.W., Palmer D.A., Chakrabarty A.M., Bender C.L. 1995; Copper as a signal for alginate synthesis in Pseudomonas syringae pv. syringae.. Appl Environ Microbiol 61:2172–2179
    [Google Scholar]
  16. Knutson C.A., Jeanes A. 1968; A new modification of the carbazole analysis: application to heteropolysaccharides.. Anal Biochem 24:470–481
    [Google Scholar]
  17. Leitão J.H., Sá-Correia I. 1993; Oxygen-dependent alginate synthesis and enzymes in Pseudomonas aeruginosa. . J Gen Microbiol 139:441–445
    [Google Scholar]
  18. Leitão J.H., Sá-Correia I. 1995; Growth-phase-dependent alginate synthesis, activity of biosynthetic enzymes and transcription of alginate genes in Pseudomonas aeruginosa. . Arch Microbiol 163:217–222
    [Google Scholar]
  19. Leitão J.H., Fialho A.M., Sá-Correia I. 1992; Effects of growth temperature on alginate synthesis and enzymes in Pseudomonas aeruginosa variants.. J Gen Microbiol 138:605–610
    [Google Scholar]
  20. McCord J.M., Fridovich I. 1969; Superoxide dismutase: an enzymatic function for erythrocuprein.. J Biol Chem 244:6049–6055
    [Google Scholar]
  21. Martin D.W., Schurr M.J., Yu H., Deretic V. 1994; Analysis of promoters controlled by the putative sigma factor AlgU regulating conversion to mucoidy in Pseudomonas aeruginosa: relationship to σ E and stress response.. J Bacteriol 176:6688–6696
    [Google Scholar]
  22. Martins L.O., Sá-Correia I. 1991; Alginate biosynthesis in mucoid recombinants of Pseudomonas aeruginosa overproducing GDP-mannose dehydrogenase.. Enzyme Microb Technol 13:385–389
    [Google Scholar]
  23. Martins L.O., Brito L.C., Sá-Correia I. 1990; Roles of Mn2+, Mg2+, and Ca2+ on alginate biosynthesis by Pseudomonas aeruginosa. . Enzyme Microb Technol 12:794–799
    [Google Scholar]
  24. May T.B., Chakrabarty A.M. 1994; Pseudomonas aeruginosa: genes and enzymes of alginate synthesis.. Trends Microbiol 2:151–157
    [Google Scholar]
  25. May T.B., Shinabarger D., Maharaj R., Kato J., Chu L, DeVault J.D., Roychoudhury S., Zielinski N.A., Berry A., Rothmel R.K., Misra T.K., Chakrabarty A.M. 1991; Alginate synthesis by Pseudomonas aeruginosa: a key pathogenic factor in chronic pulmonary infections of cystic fibrosis patients.. Clin Microbiol Rev 4:191–206
    [Google Scholar]
  26. May T.B., Shinabarger D., Boyd A., Chakrabarty A.M. 1994; Identification of amino acid residues involved in the activity of phosphomannose isomerase-guanosine 5´-diphospho-d-mannose pyrophosphorylase: a bifunctional enzyme in the alginate biosynthetic pathway of Pseudomonas aeruginosa. . J Biol Chem 269:4872–4877
    [Google Scholar]
  27. Sá-Correia I., Darzins A., Wang S.-K., Berry A., Chakrabarty A.M. 1987; Alginate biosynthetic enzymes in mucoid and nonmucoid Pseudomonas aeruginosa. Overproduction of phosphomannose isomerase, phosphomannomutase and GDP-mannose pyrophosphorylase by overexpression of the phosphomannose isomerase (pmi) gene.. J Bacteriol 169:3224–3231
    [Google Scholar]
  28. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  29. Schlictman D., Kavanaugh-Black A., Shankar S., Chakrabarty A.M. 1994; Energy metabolism and alginate biosynthesis in Pseudomonas aeruginosa: role of the tricarboxylic acid cycle.. J Bacteriol 176:6023–6029
    [Google Scholar]
  30. Schmitt M.E., Brown T.A., Trumpower B.L. 1990; A rapid and simple method for the preparation of RNA from Saccharomyces cerevisiae. . Nucleic Acids Res 18:3091
    [Google Scholar]
  31. Stock J.B., Ninfa A.J., Stock A.M. 1989; Protein phosphorylation and the regulation of adaptive responses in bacteria.. Microbiol Rev 53:450–490
    [Google Scholar]
  32. Storz G., Tartaglia L.A., Farr S.B., Ames B.N. 1990; Bacterial defenses against oxidative stress.. Trends Genet 6:363–368
    [Google Scholar]
  33. Sutherland I.W. 1990; Physiology and industrial production.. In Biotechnology of Microbial Exopolysaccharides pp. 70–88 Edited by Baddiley J., Carey N. H., Higgins I. J., Potter W. G. Cambridge: Cambridge University Press;
    [Google Scholar]
  34. Wozniak D.J., Ohman D.E. 1994; Transcriptional analysis of the Pseudomonas aeruginosa genes algR, algB, and algD reveals a hierarchy of alginate gene expression which is modulated by algT. . J Bacteriol 176:6007–6014
    [Google Scholar]
  35. Yu H., Schurr M.J., Deretic V. 1995; Functional equivalence of Escherichia coli σ E and Pseudomonas aeruginosa AlgU: E. coli ropE restores mucoidy and reduces sensitivity to reactive oxygen intermediates in algU mutants of P.aeruginosa. . J Bacteriol 177:3259–3268
    [Google Scholar]
  36. Zielinski N.A., Chakrabarty A.M., Berry A. 1991; Characterization and regulation of the Pseudomonas aeruginosa algC gene encoding phosphomannomutase.. J Biol Chem 266:9754–9763
    [Google Scholar]
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