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. 1997 Aug;179(15):4874–4881. doi: 10.1128/jb.179.15.4874-4881.1997

Osmoprotectant-dependent expression of plcH, encoding the hemolytic phospholipase C, is subject to novel catabolite repression control in Pseudomonas aeruginosa PAO1.

A E Sage 1, M L Vasil 1
PMCID: PMC179336  PMID: 9244277

Abstract

Expression of the hemolytic phospholipase C (PlcH) of Pseudomonas aeruginosa is induced under phosphate starvation conditions or in the presence of the osmoprotectants choline and glycine betaine. Because choline and glycine betaine may serve as carbon and energy sources in addition to conferring osmoprotection to P. aeruginosa, it seemed possible that induction of plcH is subject to catabolite repression control (CRC) by tricarboxylic cycle intermediates such as succinate. Total phospholipase (PLC) activity in osmoprotectant-induced cultures of P. aeruginosa PAO1 supplemented with 20 mM succinate was three- to fourfold lower than the levels in cultures supplemented with the non-catabolite-repressive substrate lactate. Analyses of osmoprotectant-dependent plcH expression in a derivative of strain PAO1 containing a plcH::lacZ operon fusion showed that (i) succinate prevented induction of plcH expression by osmoprotectants; and (ii) addition of succinate reduced or shut down further expression of plcH in osmoprotectant-induced bacteria, while cultures supplemented with lactate had little or no change in plcH expression. RNase protection analysis confirmed that repression of plcH occurs at the transcriptional level. However, a P. aeruginosa mutant decoupled in CRC exhibited a phenotype similar to that of the wild-type strain (PAO1) with respect to succinate-dependent repression of plcH expression. Osmoprotectant-induced total PLC activities, levels of expression of plcH measured with the same plcH::lacZ fusion, and levels of plcH transcription in a CRC-deficient strain reflected those seen in strain PAO1. This indicates that CRC of plcH functions by a distinct mechanism which differs from that regulating the glucose or mannitol catabolic pathway. A strain carrying a mutation in vfr, which encodes the Escherichia coli Crp homolog in P. aeruginosa, still exhibited a wild-type phenotype with respect to osmoprotectant-dependent expression and CRC of plcH. These data indicate that there is a novel CRC system that regulates the expression of plcH in P. aeruginosa.

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Selected References

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  1. Anba J., Bidaud M., Vasil M. L., Lazdunski A. Nucleotide sequence of the Pseudomonas aeruginosa phoB gene, the regulatory gene for the phosphate regulon. J Bacteriol. 1990 Aug;172(8):4685–4689. doi: 10.1128/jb.172.8.4685-4689.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berka R. M., Gray G. L., Vasil M. L. Studies of phospholipase C (heat-labile hemolysin) in Pseudomonas aeruginosa. Infect Immun. 1981 Dec;34(3):1071–1074. doi: 10.1128/iai.34.3.1071-1074.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Caminiti S. P., Young S. L. The pulmonary surfactant system. Hosp Pract (Off Ed) 1991 Jan 15;26(1):87-90, 94-100. doi: 10.1080/21548331.1991.11704128. [DOI] [PubMed] [Google Scholar]
  4. Coutinho I. R., Berk R. S., Mammen E. Platelet aggregation by a phospholipase C from Pseudomonas aeruginosa. Thromb Res. 1988 Sep 1;51(5):495–505. doi: 10.1016/0049-3848(88)90115-6. [DOI] [PubMed] [Google Scholar]
  5. Csonka L. N., Hanson A. D. Prokaryotic osmoregulation: genetics and physiology. Annu Rev Microbiol. 1991;45:569–606. doi: 10.1146/annurev.mi.45.100191.003033. [DOI] [PubMed] [Google Scholar]
  6. D'Souza-Ault M. R., Smith L. T., Smith G. M. Roles of N-acetylglutaminylglutamine amide and glycine betaine in adaptation of Pseudomonas aeruginosa to osmotic stress. Appl Environ Microbiol. 1993 Feb;59(2):473–478. doi: 10.1128/aem.59.2.473-478.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Exton J. H. Signaling through phosphatidylcholine breakdown. J Biol Chem. 1990 Jan 5;265(1):1–4. [PubMed] [Google Scholar]
  8. Gray G. L., Berka R. M., Vasil M. L. A Pseudomonas aeruginosa mutant non-derepressible for orthophosphate-regulated proteins. J Bacteriol. 1981 Aug;147(2):675–678. doi: 10.1128/jb.147.2.675-678.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gray G. L., Berka R. M., Vasil M. L. Phospholipase C regulatory mutation of Pseudomonas aeruginosa that results in constitutive synthesis of several phosphate-repressible proteins. J Bacteriol. 1982 Jun;150(3):1221–1226. doi: 10.1128/jb.150.3.1221-1226.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  11. Hancock R. E., Poole K., Benz R. Outer membrane protein P of Pseudomonas aeruginosa: regulation by phosphate deficiency and formation of small anion-specific channels in lipid bilayer membranes. J Bacteriol. 1982 May;150(2):730–738. doi: 10.1128/jb.150.2.730-738.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Holloway B. W., Krishnapillai V., Morgan A. F. Chromosomal genetics of Pseudomonas. Microbiol Rev. 1979 Mar;43(1):73–102. doi: 10.1128/mr.43.1.73-102.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johansen T., Bjørkøy G., Overvatn A., Diaz-Meco M. T., Traavik T., Moscat J. NIH 3T3 cells stably transfected with the gene encoding phosphatidylcholine-hydrolyzing phospholipase C from Bacillus cereus acquire a transformed phenotype. Mol Cell Biol. 1994 Jan;14(1):646–654. doi: 10.1128/mcb.14.1.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kurioka S., Matsuda M. Phospholipase C assay using p-nitrophenylphosphoryl-choline together with sorbitol and its application to studying the metal and detergent requirement of the enzyme. Anal Biochem. 1976 Sep;75(1):281–289. doi: 10.1016/0003-2697(76)90078-6. [DOI] [PubMed] [Google Scholar]
  15. König B., Jaeger K. E., König W. Induction of inflammatory mediator release (12-hydroxyeicosatetraenoic acid) from human platelets by Pseudomonas aeruginosa. Int Arch Allergy Immunol. 1994 May;104(1):33–41. doi: 10.1159/000236706. [DOI] [PubMed] [Google Scholar]
  16. König B., Jaeger K. E., Sage A. E., Vasil M. L., König W. Role of Pseudomonas aeruginosa lipase in inflammatory mediator release from human inflammatory effector cells (platelets, granulocytes, and monocytes. Infect Immun. 1996 Aug;64(8):3252–3258. doi: 10.1128/iai.64.8.3252-3258.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Le Rudulier D., Strom A. R., Dandekar A. M., Smith L. T., Valentine R. C. Molecular biology of osmoregulation. Science. 1984 Jun 8;224(4653):1064–1068. doi: 10.1126/science.224.4653.1064. [DOI] [PubMed] [Google Scholar]
  18. Lisa T. A., Garrido M. N., Domenech C. E. Induction of acid phosphatase and cholinesterase activities in Ps. aeruginosa and their in-vitro control by choline, acetylcholine and betaine. Mol Cell Biochem. 1983;50(2):149–155. doi: 10.1007/BF00285640. [DOI] [PubMed] [Google Scholar]
  19. Lisa T. A., Garrido M. N., Domenech C. E. Pseudomonas aeruginosa acid phosphatase and cholinesterase induced by choline and its metabolic derivatives may contain a similar anionic peripheral site. Mol Cell Biochem. 1984 Sep;63(2):113–118. doi: 10.1007/BF00285217. [DOI] [PubMed] [Google Scholar]
  20. Lucchesi G. I., Lisa T. A., Domenech C. E. Choline and betaine as inducer agents of Pseudomonas aeruginosa phospholipase C activity in high phosphate medium. FEMS Microbiol Lett. 1989 Feb;57(3):335–338. doi: 10.1016/0378-1097(89)90324-8. [DOI] [PubMed] [Google Scholar]
  21. Meyers D. J., Berk R. S. Characterization of phospholipase C from Pseudomonas aeruginosa as a potent inflammatory agent. Infect Immun. 1990 Mar;58(3):659–666. doi: 10.1128/iai.58.3.659-666.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ng F. M., Dawes E. A. Chemostat studies on the regulation of glucose metabolism in Pseudomonas aeruginosa by citrate. Biochem J. 1973 Feb;132(2):129–140. doi: 10.1042/bj1320129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ng F. M., Dawes E. A. Regulation of enzymes of glucose metabolism by citrate in Pseudomonas aeruginosa. Biochem J. 1967 Sep;104(3):48P–48P. [PMC free article] [PubMed] [Google Scholar]
  24. Ostroff R. M., Vasil A. I., Vasil M. L. Molecular comparison of a nonhemolytic and a hemolytic phospholipase C from Pseudomonas aeruginosa. J Bacteriol. 1990 Oct;172(10):5915–5923. doi: 10.1128/jb.172.10.5915-5923.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ostroff R. M., Vasil M. L. Identification of a new phospholipase C activity by analysis of an insertional mutation in the hemolytic phospholipase C structural gene of Pseudomonas aeruginosa. J Bacteriol. 1987 Oct;169(10):4597–4601. doi: 10.1128/jb.169.10.4597-4601.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ostroff R. M., Wretlind B., Vasil M. L. Mutations in the hemolytic-phospholipase C operon result in decreased virulence of Pseudomonas aeruginosa PAO1 grown under phosphate-limiting conditions. Infect Immun. 1989 May;57(5):1369–1373. doi: 10.1128/iai.57.5.1369-1373.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pritchard A. E., Vasil M. L. Nucleotide sequence and expression of a phosphate-regulated gene encoding a secreted hemolysin of Pseudomonas aeruginosa. J Bacteriol. 1986 Jul;167(1):291–298. doi: 10.1128/jb.167.1.291-298.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rahme L. G., Stevens E. J., Wolfort S. F., Shao J., Tompkins R. G., Ausubel F. M. Common virulence factors for bacterial pathogenicity in plants and animals. Science. 1995 Jun 30;268(5219):1899–1902. doi: 10.1126/science.7604262. [DOI] [PubMed] [Google Scholar]
  29. Saiman L., Cacalano G., Gruenert D., Prince A. Comparison of adherence of Pseudomonas aeruginosa to respiratory epithelial cells from cystic fibrosis patients and healthy subjects. Infect Immun. 1992 Jul;60(7):2808–2814. doi: 10.1128/iai.60.7.2808-2814.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saiman L., Ishimoto K., Lory S., Prince A. The effect of piliation and exoproduct expression on the adherence of Pseudomonas aeruginosa to respiratory epithelial monolayers. J Infect Dis. 1990 Mar;161(3):541–548. doi: 10.1093/infdis/161.3.541. [DOI] [PubMed] [Google Scholar]
  31. Salvano M. A., Lisa T. A., Domenech C. E. Choline transport in Pseudomonas aeruginosa. Mol Cell Biochem. 1989 Jan 23;85(1):81–89. doi: 10.1007/BF00223517. [DOI] [PubMed] [Google Scholar]
  32. Shortridge V. D., Lazdunski A., Vasil M. L. Osmoprotectants and phosphate regulate expression of phospholipase C in Pseudomonas aeruginosa. Mol Microbiol. 1992 Apr;6(7):863–871. doi: 10.1111/j.1365-2958.1992.tb01537.x. [DOI] [PubMed] [Google Scholar]
  33. West S. E., Sample A. K., Runyen-Janecky L. J. The vfr gene product, required for Pseudomonas aeruginosa exotoxin A and protease production, belongs to the cyclic AMP receptor protein family. J Bacteriol. 1994 Dec;176(24):7532–7542. doi: 10.1128/jb.176.24.7532-7542.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolff J. A., MacGregor C. H., Eisenberg R. C., Phibbs P. V., Jr Isolation and characterization of catabolite repression control mutants of Pseudomonas aeruginosa PAO. J Bacteriol. 1991 Aug;173(15):4700–4706. doi: 10.1128/jb.173.15.4700-4706.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wood R. E. Pseudomonas: the compromised host. Hosp Pract. 1976 Aug;11(8):91–100. doi: 10.1080/21548331.1976.11706983. [DOI] [PubMed] [Google Scholar]
  36. Zach M. S. Lung disease in cystic fibrosis--an updated concept. Pediatr Pulmonol. 1990;8(3):188–202. doi: 10.1002/ppul.1950080311. [DOI] [PubMed] [Google Scholar]
  37. von Gabain A., Belasco J. G., Schottel J. L., Chang A. C., Cohen S. N. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]

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