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Volume 154, Issue 6

The PsrA responds to long-chain fatty acid signals to regulate the -oxidation operon Free

Abstract

-Oxidative enzymes for fatty acid degradation (Fad) of long-chain fatty acids (LCFAs) are induced during lung infection in cystic fibrosis patients, and this may contribute to nutrient acquisition and pathogenesis of . The promoter region of one -oxidation operon, (PA3014 and PA3013), was mapped. Focusing on the transposon mutagenesis of strain PAO1 carrying the P fusion, a regulator for the operon was identified to be PsrA (PA3006). Transcriptome analysis of the Δ mutant indicated its importance in regulating -oxidative enzymes. These microarray data were confirmed by real-time RT-PCR analyses of the and (encoding a lipase) genes. Induction of the operon was demonstrated to respond to novel LCFA signals, and this induction required the presence of PsrA, suggesting that LCFAs bind to PsrA to derepress . Electrophoretic mobility shift assays indicate specific binding of PsrA to the promoter region. This binding is disrupted by specific LCFAs (C , C, C and, to a lesser extent, C), but not by other medium- or short-chain fatty acids or the first intermediate of -oxidation, acyl-CoA. It is shown here that PsrA is a regulator that binds and responds to LCFA signals in .

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  • Accepted:
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Keyword(s): CF, cystic fibrosis , EMSA, electrophoretic mobility shift assay , FA, fatty acid , Fad, fatty acid degradation , LCFA, long-chain fatty acid , MCFA, medium-chain fatty acid , PC, phosphatidylcholine and SCFA, short-chain fatty acid
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2008-06-01
2024-04-16
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References

  1. Baltch A. L., Griffin P. E. 1977; Pseudomonas aeruginosa bacteremia: clinical study of 75 patients. Am J Med Sci 274:119–129
     [Google Scholar]
  2. Bowton D. L. 1999; Nosocomial pneumonia in the ICU – year 2000 and beyond. Chest 115:3 Suppl.28S–33S
     [Google Scholar]
  3. Campbell J. W., Cronan J. E. Jr 2002; The enigmatic Escherichia coli fadE gene is yafH . J Bacteriol 184:3759–3764
     [Google Scholar]
  4. Chen C.-H., Cheng J.-C., Cho Y.-C., Hsu W.-H. 2005; A gene cluster for the fatty acid catabolism from Pseudonocardia autotrophica BCRC12444. Biochem Biophys Res Commun 329:863–868
     [Google Scholar]
  5. Choi K.-H., Kumar A., Schweizer H. P. 2006; A 10-min method for preparation of highly electrocompetent Pseudomonas aeruginosa cells: application for DNA fragment transfer between chromosomes and plasmid transformation. J Microbiol Methods 64:391–397
     [Google Scholar]
  6. Cronan J. E. Jr, Satyanarayana S. 1998; FadR, transcriptional co-ordination of metabolic expediency. Mol Microbiol 29:937–943
     [Google Scholar]
  7. Cronin C. N., McIntire W. S. 1999; pUCP-Nco and pUCP-Nde: Escherichia–Pseudomonas shuttle vectors for recombinant protein expression in Pseudomonas . Anal Biochem 272:112–115
     [Google Scholar]
  8. DiRusso C. C., Heimert T. L., Metzger A. K. 1992; Characterization of FadR, a global transcription regulator of fatty acid metabolism in Escherichia coli . J Biol Chem 267:8685–8691
     [Google Scholar]
  9. Doring G. 1997; Cystic fibrosis respiratory infections: interactions between bacteria and host defense. Monaldi Arch Chest Dis 52:363–366
     [Google Scholar]
  10. Fleiszig S. M. J., Zaidl T. S., Pier G. B. 1995; Pseudomonas aeruginosa invasion of and multiplication within corneal epithelial cells in vitro. Infect Immun 63:4072–4077
     [Google Scholar]
  11. Gordon L., Chervonenkis A. Y., Gammerman A. J., Shahmuradov I. A., Solovyev V. V. 2003; Sequence alignment kernel for recognition of promoter regions. Bioinformatics 19:1964–1971
     [Google Scholar]
  12. Greenberger P. A. 1997; Immunologic aspects of lung diseases and cystic fibrosis. JAMA 278:1924–1930
     [Google Scholar]
  13. Henry M. F., Cronan J. E. 1992; A new mechanism of transcriptional regulation: release of an activator triggered by small molecule binding. Cell 70:671–679
     [Google Scholar]
  14. Hoang T. T., Karkhoff-Schweizer R. R., Kutchma A. J., Schweizer H. P. 1998; A broad-host-range Flp- FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 212:77–86
     [Google Scholar]
  15. Hoang T. T., Kutchma A. J., Becher A., Schweizer H. P. 2000; Integration-proficient plasmids for Pseudomonas aeruginosa : site-specific integration and use for engineering of reporter and expression strains. Plasmid 43:59–72
     [Google Scholar]
  16. Hoang T. T., Sullivan S. A., Cusick K. C., Schweizer P. H. 2002; β -Ketoacyl carrier protein reductase (FabG) activity of the fatty acid biosynthetic pathway is a determining factor of 3-oxo-homoserine lactone acyl chain lengths. Microbiology 148:3849–3856
     [Google Scholar]
  17. Holloway B. W., Roemling U., Tuemmler B. 1994; Genomic mapping of Pseudomonas aeruginosa PAO. Microbiology 140:2907–2929
     [Google Scholar]
  18. Kojic M., Aguilar C., Venturi V. 2002; TetR family member PsrA directly binds the Pseudomonas rpoS and psrA promoters. J Bacteriol 184:2324–2330
     [Google Scholar]
  19. Kojic M., Jovcic B., Vindigni A., Odreman F., Venturi V. 2005; Novel target genes of PsrA transcriptional regulator of Pseudomonas aeruginosa . FEMS Microbiol Lett 246:175–181
     [Google Scholar]
  20. Kulasekara H. D., Ventre I., Kulasekara B. R., Lazdunski A., Filloux A., Lory S. 2005; A novel two-component system controls the expression of Pseudomonas aeruginosa fimbrial cup genes. Mol Microbiol 55:368–380
     [Google Scholar]
  21. Levano-Garcia J., Verjovski-Almeida S., da Silva A. C. R. 2005; Mapping transposon insertion sites by touchdown PCR and hybrid degenerate primers. Biotechniques 38:225–229
     [Google Scholar]
  22. Lode H., Raffenberg M., Erbes R., Geerdes-Fenge H., Mauch H. 2000; Nosocomial pneumonia: epidemiology, pathogenesis, diagnosis, treatment and prevention. Curr Opin Infect Dis 13:377–384
     [Google Scholar]
  23. Martinez A., Ostrovsky P., Nunn D. N. 1999; LipC, a second lipase of Pseudomonas aeruginosa , is LipB and Xcp dependent and is transcriptionally regulated by pilus biogenesis components. Mol Microbiol 34:317–326
     [Google Scholar]
  24. Miller J. H. 1992 A Short Course in Bacterial Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  25. Pennington J. E. 1994; Pseudomonas aeruginosa pneumonia and other respiratory tract infections. In Pseudomonas aeruginosa: Infections and Treatment pp 159–181 Edited by Baltch A. L., Smith R. P. New York: Marcel Dekker;
     [Google Scholar]
  26. Postle A. D., Mander A., Reid K. B. M., Wang J.-Y., Wright S. M., Moustaki M., Warner J. O. 1999; Deficient hydrophilic lung surfactant proteins A and D with normal surfactant phospholipid molecular species in cystic fibrosis. Am J Respir Cell Mol Biol 20:90–98
     [Google Scholar]
  27. Pruitt B. A. Jr, McManus A. T., Kim S. H., Goodwin C. W. 1998; Burn wound infections: current status. World J Surg 22:135–145
     [Google Scholar]
  28. Reyes M. P., Lerner A. M. 1983; Current problems in the treatment of infective endocarditis due to P. aeruginosa . Rev Infect Dis 5:314–321
     [Google Scholar]
  29. Richards M. J., Edwards J. R., Culver D. H., Gaynes R. P. 1999; Nosocomial infections in medical intensive care units in the United States. Crit Care Med 27:887–892
     [Google Scholar]
  30. Rosenau F., Jaeger K.-E. 2000; Bacterial lipases from Pseudomonas : regulation of gene expression and mechanisms of secretion. Biochimie 82:1023–1032
     [Google Scholar]
  31. Salaun J. P., Helvig C. 1995; Cytochrome P450 dependent oxidation of fatty acids. Drug Metabol Drug Interact 12:261–283
     [Google Scholar]
  32. Sambrook J., Russell D. W. 2001 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  33. Schaberg D. R., Culver D. H., Gaynes R. P. 1991; Major trends in the microbial etiology of nosocomial infection. Am J Med 91 :suppl. 3B72S–75S
     [Google Scholar]
  34. Schweizer H. P. 1994; A method for construction of bacterial hosts for lac -based cloning and expression vectors: α -complementation and regulated expression. Biotechniques 17:452–456
     [Google Scholar]
  35. Schweizer H. P., Chuanshuen R. 2001; A small broad-host-range lacZ operon fusion vector with low background activity. Biotechniques 31:1258–1262
     [Google Scholar]
  36. Shen D. K., Filopon D., Kuhn L., Polack B., Toussaint B. 2006; PsrA is a positive transcriptional regulator of the type III secretion system in Pseudomonas aeruginosa . Infect Immun 74:1121–1129
     [Google Scholar]
  37. Son M. S., Matthews W. J. Jr, Kang Y., Nguyen D. T., Hoang T. T. 2007; In vivo evidence of Pseudomonas aeruginosa nutrient acquisition and pathogenesis in the lungs of cystic fibrosis patients. Infect Immun 75:5313–5324
     [Google Scholar]
  38. Son M. S., Nguyen D. T., Kang Y., Hoang T. T. 2008; Engineering of FRT–lacZ fusion constructs: induction of the Pseudomonas aeruginosa fadBA1 operon by medium and long chain-length fatty acids. Plasmid 59:111–118
     [Google Scholar]
  39. Stover C. K., Pham X. Q., Erwin A. L., Mizoguchi S. D., Warrener P., Hickey M. J., Brinkman F. S., Hufnagle W. O. other authors 2000; Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406:959–964
     [Google Scholar]
  40. Tabansky I., Nurminsky D. I. 2003; Mapping of transcription start sites by direct sequencing of SMARTTM . RACE products. Biotechniques 34:482–486
     [Google Scholar]
  41. Vandesompele J., De Preter K., Pattyn F., Poppe B., Van Roy N., De Paepe A., Speleman F. 2002; Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3: research0034
    [Google Scholar]
  42. Yang X. Y., Schulz H., Elzinga M., Yang S. Y. 1991; Nucleotide sequence of the promoter and fadB gene of the fadBA operon and primary structure of the multifunctional fatty acid oxidation protein from Escherichia coli . Biochemistry 30:6788–6795
     [Google Scholar]
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The Pseudomonas aeruginosa PsrA responds to long-chain fatty acid signals to regulate the fadBA5β-oxidation operon
Microbiology 154, 1584 (2008); https://doi.org/10.1099/mic.0.2008/018135-0
/content/journal/micro/10.1099/mic.0.2008/018135-0
/content/journal/micro/10.1099/mic.0.2008/018135-0
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