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. 1984 Jun;158(3):801–808. doi: 10.1128/jb.158.3.801-808.1984

Genetic mapping and characterization of Pseudomonas aeruginosa mutants defective in the formation of extracellular proteins.

B Wretlind, O R Pavlovskis
PMCID: PMC215512  PMID: 6427194

Abstract

We isolated 15 mutants of Pseudomonas aeruginosa PAO which were defective in the formation of certain extracellular proteins, such as elastase, staphylolytic enzyme, and lipase ( Xcp mutants). The mutations were mapped on the chromosome by conjugation and transduction. The locations were xcp -1 near 0', with the gene order cys-59- xcp -1- proB , and loci xcp -2, xcp -3, and xcp -31 at 35', with the gene order trpC , D- xcp -3/ xcp -31- xcp -2- argC . Loci xcp -4 and xcp -41 through xcp -44 were cotransducible with proA at 40'; loci xcp -5, xcp -51, xcp -52, and xcp53 were located at 55', with the gene order leu-10- trpF -met-9010- xcp -53- xcp -5/ xcp -51/ xcp+ ++-52, and xcp -6 was located at 65' to 70', between catA and mtu-9002. Nine mutations ( xcp -2, xcp -3, xcp -31, xcp -4, and xcp -41 through xcp -45) caused decreased production of extracellular enzymes. Six strains with mutations xcp -1, xcp -5, xcp -51, xcp -52, xcp -53, and xcp -6 produced cell-bound exoproteins and had defective release mechanisms. The regulation of production of alkaline phosphatase and phospholipase C is different from other exoproteins , such as elastase, but they all seem to share a common release mechanism. Alkaline protease had separate mechanisms for regulation and release, since this protease was found in culture supernatants of all but one of the mutants, and none of the strains had cell-bound enzyme.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. Bjorn M. J., Sokol P. A., Iglewski B. H. Influence of iron on yields of extracellular products in Pseudomonas aeruginosa cultures. J Bacteriol. 1979 Apr;138(1):193–200. doi: 10.1128/jb.138.1.193-200.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Datta N., Hedges R. W., Shaw E. J., Sykes R. B., Richmond M. H. Properties of an R factor from Pseudomonas aeruginosa. J Bacteriol. 1971 Dec;108(3):1244–1249. doi: 10.1128/jb.108.3.1244-1249.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeBell R. M. Production of exotoxin A by Pseudomonas aeruginosa in a chemically defined medium. Infect Immun. 1979 Apr;24(1):132–138. doi: 10.1128/iai.24.1.132-138.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glenn A. R. Production of extracellular proteins by bacteria. Annu Rev Microbiol. 1976;30:41–62. doi: 10.1146/annurev.mi.30.100176.000353. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Gray G. L., Vasil M. L. Isolation and genetic characterization of toxin-deficient mutants of Pseudomonas aeruginosa PAO. J Bacteriol. 1981 Aug;147(2):275–281. doi: 10.1128/jb.147.2.275-281.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Haas D., Holloway B. W. Chromosome mobilization by the R plasmid R68.45: a tool in Pseudomonas genetics. Mol Gen Genet. 1978 Jan 17;158(3):229–237. doi: 10.1007/BF00267194. [DOI] [PubMed] [Google Scholar]
  10. Haas D., Holloway B. W. R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet. 1976 Mar 30;144(3):243–251. doi: 10.1007/BF00341722. [DOI] [PubMed] [Google Scholar]
  11. Haas D., Holloway B. W., Schamböck A., Leisinger T. The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet. 1977 Jul 7;154(1):7–22. doi: 10.1007/BF00265571. [DOI] [PubMed] [Google Scholar]
  12. Hanne L. F., Howe T. R., Iglewski B. H. Locus of the Pseudomonas aeruginosa toxin A gene. J Bacteriol. 1983 Apr;154(1):383–386. doi: 10.1128/jb.154.1.383-386.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Howard S. P., Buckley J. T. Intracellular accumulation of extracellular proteins by pleiotropic export mutants of Aeromonas hydrophila. J Bacteriol. 1983 Apr;154(1):413–418. doi: 10.1128/jb.154.1.413-418.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Howe T. R., Wretlind B., Iglewski B. H. Comparison of two methods of genetic exchange in determination of the genetic locus of the structural gene for Pseudomonas aeruginosa elastase. J Bacteriol. 1983 Oct;156(1):58–61. doi: 10.1128/jb.156.1.58-61.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Iglewski B. H., Kabat D. NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin,. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2284–2288. doi: 10.1073/pnas.72.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Isaac J. H., Holloway B. W. Control of arginine biosynthesis in Pseudomonas aeruginosa. J Gen Microbiol. 1972 Dec;73(3):427–438. doi: 10.1099/00221287-73-3-427. [DOI] [PubMed] [Google Scholar]
  18. Isaac J. H., Holloway B. W. Control of pyrimidine biosynthesis in Pseudomonas aeruginosa. J Bacteriol. 1968 Nov;96(5):1732–1741. doi: 10.1128/jb.96.5.1732-1741.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jensen S. E., Fecycz I. T., Campbell J. N. Nutritional factors controlling exocellular protease production by Pseudomonas aeruginosa. J Bacteriol. 1980 Nov;144(2):844–847. doi: 10.1128/jb.144.2.844-847.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Krishnapillai V. A novel transducing phage. Its role in recognition of a possible new host-controlled modification system in Pseudomonas aeruginosa. Mol Gen Genet. 1972;114(2):134–143. doi: 10.1007/BF00332784. [DOI] [PubMed] [Google Scholar]
  21. Krishnapillai V., Royle P., Lehrer J. Insertions of the transposon Tn1 into the Pseudomonas aeruginosa chromosome. Genetics. 1981 Mar-Apr;97(3-4):495–511. doi: 10.1093/genetics/97.3-4.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lindström E. B. Alternative replica plating technique. Appl Environ Microbiol. 1977 Aug;34(2):225–227. doi: 10.1128/aem.34.2.225-227.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Liu P. V. Exotoxins of Pseudomonas aeruginosa. I. Factors that influence the production of exotoxin A. J Infect Dis. 1973 Oct;128(4):506–513. doi: 10.1093/infdis/128.4.506. [DOI] [PubMed] [Google Scholar]
  24. Lory S., Tai P. C., Davis B. D. Mechanism of protein excretion by gram-negative bacteria: Pseudomonas aeruginosa exotoxin A. J Bacteriol. 1983 Nov;156(2):695–702. doi: 10.1128/jb.156.2.695-702.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. MORIHARA K. PRODUCTION OF ELASTASE AND PROTEINASE BY PSEUDOMONAS AERUGINOSA. J Bacteriol. 1964 Sep;88:745–757. doi: 10.1128/jb.88.3.745-757.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. MORIHARA K., TSUZUKI H., OKA T., INOUE H., EBATA M. PSEUDOMONAS AERUGINOSA ELASTASE. ISOLATION, CRYSTALLIZATION, AND PRELIMINARY CHARACTERIZATION. J Biol Chem. 1965 Aug;240:3295–3304. [PubMed] [Google Scholar]
  27. Moore A. T., Nayudu M., Holloway B. W. Genetic mapping in Methylophilus methylotrophus AS1. J Gen Microbiol. 1983 Mar;129(3):785–799. doi: 10.1099/00221287-129-3-785. [DOI] [PubMed] [Google Scholar]
  28. Morgan A. F. Isolation and characterization of Pseudomonas aeruginosa R' plasmids constructed by interspecific mating. J Bacteriol. 1982 Feb;149(2):654–661. doi: 10.1128/jb.149.2.654-661.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Morihara K., Tsuzuki H. Production of protease and elastase by Pseudomonas aeruginosa strains isolated from patients. Infect Immun. 1977 Mar;15(3):679–685. doi: 10.1128/iai.15.3.679-685.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Neill R. J., Ivins B. E., Holmes R. K. Synthesis and secretion of the plasmid-coded heat-labile enterotoxin of Escherichia coli in Vibrio cholerae. Science. 1983 Jul 15;221(4607):289–291. doi: 10.1126/science.6857285. [DOI] [PubMed] [Google Scholar]
  31. Nielsen J. B., Mézes P. S., Lampen J. O. Secretion by Pseudomonas aeruginosa: fate of a cloned gram-positive lipoprotein deletion mutant. J Bacteriol. 1983 Nov;156(2):559–566. doi: 10.1128/jb.156.2.559-566.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pemberton J. M., Holloway B. W. A new sex factor of Pseudomonas aeruginosa. Genet Res. 1973 Jun;21(3):263–272. doi: 10.1017/s0016672300013458. [DOI] [PubMed] [Google Scholar]
  33. Pemberton J. M., Holloway B. W. Chromosome mapping in Pseudomonas aeruginosa. Genet Res. 1972 Jun;19(3):251–260. doi: 10.1017/s0016672300014518. [DOI] [PubMed] [Google Scholar]
  34. Rella M., Haas D. Resistance of Pseudomonas aeruginosa PAO to nalidixic acid and low levels of beta-lactam antibiotics: mapping of chromosomal genes. Antimicrob Agents Chemother. 1982 Aug;22(2):242–249. doi: 10.1128/aac.22.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Royle P. L., Matsumoto H., Holloway B. W. Genetic circularity of the Pseudomonas aeruginosa PAO chromosome. J Bacteriol. 1981 Jan;145(1):145–155. doi: 10.1128/jb.145.1.145-155.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Silhavy T. J., Benson S. A., Emr S. D. Mechanisms of protein localization. Microbiol Rev. 1983 Sep;47(3):313–344. doi: 10.1128/mr.47.3.313-344.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Soldati L., Crockett R., Carrigan J. M., Leisinger T., Holloway B. W., Haas D. Revised locations of the hisI and pru (proline utilization) genes on the Pseudomonas aeruginosa chromosome map. Mol Gen Genet. 1984;193(3):431–436. doi: 10.1007/BF00382080. [DOI] [PubMed] [Google Scholar]
  38. Stanisich V. A., Holloway B. W. A mutant sex factor of Pseudomonas aeruginosa. Genet Res. 1972 Feb;19(1):91–108. doi: 10.1017/s0016672300014294. [DOI] [PubMed] [Google Scholar]
  39. Stanisich V., Holloway B. W. Conjugation in Pseudomonas aeruginosa. Genetics. 1969 Feb;61(2):327–339. doi: 10.1093/genetics/61.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vasil M. L., Berka R. M., Gray G. L., Nakai H. Cloning of a phosphate-regulated hemolysin gene (phospholipase C) from Pseudomonas aeruginosa. J Bacteriol. 1982 Oct;152(1):431–440. doi: 10.1128/jb.152.1.431-440.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Vesterberg O. Quantification of proteins with a new sensitive method--zone immunoelectrophoresis assay. Hoppe Seylers Z Physiol Chem. 1980 May;361(5):617–624. doi: 10.1515/bchm2.1980.361.1.617. [DOI] [PubMed] [Google Scholar]
  42. Watson J. M., Holloway B. W. Suppressor mutations in Pseudomonas aeruginosa. J Bacteriol. 1976 Mar;125(3):780–786. doi: 10.1128/jb.125.3.780-786.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wretlind B., Hedén L., Sjöberg L., Wadström T. Production of enzymes and toxins by hospital strains of Pseudomonas aeruginosa in relation to serotype and phage-typing pattern. J Med Microbiol. 1973 Feb;6(1):91–100. doi: 10.1099/00222615-6-1-91. [DOI] [PubMed] [Google Scholar]
  44. Wretlind B., Sjöberg L., Wadström T. Protease-deficient mutants of Pseudomonas aeruginosa: pleiotropic changes in activity of other extracellular enzymes. J Gen Microbiol. 1977 Dec;103(2):329–336. doi: 10.1099/00221287-103-2-329. [DOI] [PubMed] [Google Scholar]
  45. Wretlind B., Wadström T. Purification and properties of a protease with elastase activity from Pseudomonas aeruginosa. J Gen Microbiol. 1977 Dec;103(2):319–327. doi: 10.1099/00221287-103-2-319. [DOI] [PubMed] [Google Scholar]

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