Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Multiple outer membrane receptors for uptake of ferric pseudobactins inPseudomonas putida WCS358

  • 53 Accesses

  • 27 Citations


Under iron limitationPseudomonas putida WCS358 produces a fluorescent siderophore, pseudobactin 358, which, after complexing iron, is transported back into the cell via the specific outer membrane receptor PupA. In addition, this strain has the capacity to take up iron via a large variety of siderophores produced by other fluorescent pseudomonads. Putative receptor genes for such siderophores were identified in the chromosome of strain WCS358 by PCR using primers matching two domains conserved in four ferric pseudobactin receptors, including PupA. Eleven amplification products within the expected size range were obtained. Sequence analysis confirmed that the products were derived from genes encoding outer membrane receptors. Two complete receptor genes were isolated from a genomic library ofP. putida WCS358. Both protein products are involved in the transport of a limited number of specific ferric pseudobactins. These results indicate that the ability ofP. putida WCS358 to exploit many different heterologous pseudobactins is related to the presence of multiple outer membrane receptor proteins.

This is a preview of subscription content, log in to check access.


  1. Abdallah MA (1991) Pyoverdins and pseudobactins. In: Winkelmann G (ed) CRC Handbook of Microbial Iron Chelates. CRC Press, Boca Raton, pp 139–153

  2. Bakker PAHM, Lamers JG, Bakker AW, Marugg JD, Weisbeek PJ, Schippers B (1986) The role of siderophores in potato tuber yield increase byPseudomonas putida in a short rotation of potato. Neth J Plant Pathol 92:249–256

  3. Bakker PAHM, van Peer R, Schippers B (1990) Specificity of siderophores and siderophore receptors and biocontrol byPseudomonas spp. In: Hornby D (ed) Biological control of soilborne plant pathogens. CAB International, Wallingford, UK, pp 131–142

  4. Beringer JE, Beynon JL, Buchanan-Wollaston AV, Johnston AWB (1978) Transfer of the drug-resistance transposon Tn5 toRhizobium. Nature 276:633–634

  5. Better M, Lewis B, Corbin D, Ditta G, Helinski DR (1983) Structural relationships amongRhizobium meliloti promoters. Cell 35:479–485

  6. Birnboim HC (1983) A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol 100:243–255

  7. Bitter W, Marugg JD, de Weger LA, Tommassen J, Weisbeek PJ (1991) The ferric-pseudobactin receptor PupA ofPseudomonas putida WCS358: homology to TonB-dependentEscherichia coli receptors and specifity of the protein. Mol Microbial 5:647–655

  8. Bitter W, Tommassen J, Weisbeek PJ (1993) Identification and characterization of theexbB, exbD andtonB genes ofPseudomonas putida WCS358: their involvement in ferric-pseudobactin transport. Mol Microbiol 7:117–130

  9. Bonas U, Stall RE, Staskawicz B (1989) Genetic and structural characterization of the avirulence geneavrBs3 fromXanthomonas campestris pv.vesicatoria. Mol Gen Genet 218:127–136

  10. Buyer JS, Sikora LJ, Chaney RL (1989) A new growth medium for the study of siderophore-mediated interactions. Biol Fertil Soils 8:98–101

  11. Cohen SN, Chang ACY, Hsu CL (1972) Nonchromosomal antibiotic resistance in bacteria; genetic transformation ofEscherichia coli by R factor DNA. Proc Natl Acad Sci USA 69:2110–2114

  12. Coulton JW, Mason P, Cameron DR, Carmel G, Jean R, Rode HN (1986) Protein fusions ofβ-galactosidase to the ferrichrome-iron receptor ofEscherichia coli K-12. J Bacteriol 165:181–192

  13. Dean CR, Poole K (1993) Cloning and characterization of the ferric enterobactin receptor gene (pfeA) ofPseudomonas aeruginosa. J Bacteriol 175:317–324

  14. De Weger LA, van Boxtel R, van der Burg B, Gruters R, Geels FP, Schippers B, Lugtenberg B (1986) Siderophores and outer membrane proteins of antagonistic, plant-growth stimulating, rootcolonizingPseudomonas spp. J Bacteriol 165:585–594

  15. Dente L, Cesareni G, Cortese R (1983) pEMBL: A new family of single stranded plasmids. Nucl Acids Res 11:1645–1655

  16. Doudoroff M, Palleroni NJ (1974) Genus Pseudomonas. In: Buchanon RE, Gibbons NE (eds) Bergey's Manual of Determinative Bacteriology. Williams and Wilkins, Baltimore, Md., pp 217–243

  17. Figurski DH, Helinski DR (1979) Replication of an origin containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

  18. Geels FP, Schippers B (1983) Reduction in yield depressions in high frequency potato cropping soil after seed tuber treatments with antognistic fluorescentPseudomonas spp. Phytopathol Z 108:207–221

  19. Haas D, Holloway BW (1976) R factor variant with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet 144:243–251

  20. Higgins TG, Sharp PM (1988) CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene 73:237–244

  21. Hohnadel D, Meyer J (1988) Specificity of pyoverdine-mediated iron uptake among fluorescentPseudomonas strains. J Bacteriol 170:4865–4873

  22. Jurkevitch E, Hadar Y, Chen Y, Libman J, Shanzer A (1992) Iron uptake and molecular recognition inPseudomonas putida: receptor mapping with ferrichrome and its biomimetic analogs. J Bacteriol 174-78:83

  23. King EO, Ward MK, Raney DE (1954) Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med 44:301–307

  24. Koebnik R, Hantke K, Braun V (1993) The TonB-dependent ferrichrome receptor FcuA ofYersinia enterocolitica: evidence against a strict co-evolution of receptor structure and substrate specificity. Mol Microbiol 7:383–393

  25. Koster M, van de Vossenberg J, Leong J, Weisbeek PJ (1993) Identification and characterization of thepupB gene encoding an inducible ferric-pseudobactin receptor ofPseudomonas putida WCS358. Mol Microbiol 8:591–601

  26. Koster M, van Klompenburg W, Bitter W, Leong J, Weisbeek P (1994) Role for the outer membrane ferric siderophore receptor PupB in signal transduction across the bacterial cell envelope. EMBO J 13:2805–2813

  27. Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685

  28. Magazin M, Moores JC, Leong J (1986) Cloning of the gene coding for the outer membrane receptor protein for ferric-pseudobactin, a siderophore from a plant growth-promotingPseudomonas strain. J Biol Chem 261:795–799

  29. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  30. Marugg JD, de Weger LA, Nielander HB, Oorthuizen M, Recourt K, Lugtenberg B, van der Hofstad GAJM, Weisbeek PJ (1989) Cloning and characterization of a gene encoding an outer membrane protein required for siderophore mediated uptake of Fe3+ inPseudomonas putida WCS358. J Bacteriol 171:2819–2826

  31. Marugg JD, Nielander HB, Horrevoets AJG, van Megen I, van Genderen I, Weisbeek PJ (1988) Genetic organization and transcriptional analysis of a major gene cluster involved in siderophore biosynthesis inPseudomonas putida WCS358. J Bacteriol 170:1812–1819

  32. Miller JH (1972) Experiments in molecular biology. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  33. Morris J, Donnelly DF, O'Neill E, McConnell F, O'Gara F (1994) Nucleotide sequence analysis and potential environment distribution of a ferric pseudobactin receptor gene ofPseudomonas sp. strain M114. Mol Gen Genet 242:9–16

  34. Neilands JB (1981) Microbial iron compounds. Annu Rev Biochem 50:715–731

  35. Neilands JB (1982) Microbial envelope proteins related to iron. Annu Rev Microbiol 36:285–309

  36. Poole K, Neshat S, Krebes K, Heinrichs DE (1993) Cloning and nucleotide sequence analysis of the ferripyoverdine receptor genefpvA ofPseudomonas aeruginosa. J Bacteriol 175:4597–4604

  37. Postle K (1990) TonB and the Gram-negative dilemma. Mol Microbiol 4:2019–2025

  38. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inibitors. Proc Natl Acad Sci USA 74:5463–5467

  39. Sauer M, Hantke K, Braun V (1990) Sequence of thefhuE outer membrane receptor gene ofEscherichia coli K12 and properties of mutants. Mol Microbiol 4:427–437

  40. Stachel SE, An G, Flores C, Nester EW (1985) A Tn3lacZ transposon for the random generation ofβ-galactosidase gene fusions: application to the analysis of gene expression inAgrobacterium. EMBO J 4:891–898

  41. Staskawicz B, Dahlbeck D, Keen N, Napoli C (1987) Molecular characterization of cloned avirulence genes from race 0 and race 1 ofPseudomonas syringae pv.glycinea. J Bacteriol 169:5789–5794

  42. Suslow TV, Schroth MN (1982) Role of deleterious rhizobacteria as minor pathogens in reducing crop growth. Phytopathology 72:111–115

  43. Tommassen J (1988) Biogenesis and membrane topology of outer membrane proteins inEscherichia coli. In: Op den Kamp JAF (ed) Membrane biogenesis. Springer-Verlag, Berlin-Heidelberg, pp 351–373

  44. Van der Hofstad GAJM, Marugg JD, Verjans GMGM, Weisbeek PJ (1986) Characterization and structural analysis of the siderophore produced by the PGPR Pseudomonas putida strain WCS358. In: Swinburne TR (ed) Iron, siderophores, and plant diseases. Plenum Press, New York, pp 71–75

  45. Wendenbaum S, Demange P, Dell A, Meyer J, Abdallah MA (1983) The structure of pyoverdinepa, the siderophore ofPseudomonas aeruginosa. Tetrahedron Lett 24:4877–4880

  46. Yang C, Leong J (1984) Structure of pseudobactin 7SR1, a siderophore from a plant deleteriousPseudomonas. Biochemistry 23:3534–3540

Download references

Author information

Correspondence to Margot Koster.

Additional information

Communicated by A. Kondorosi

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ovaa, W., Bitter, W., Weisbeek, P. et al. Multiple outer membrane receptors for uptake of ferric pseudobactins inPseudomonas putida WCS358. Molec. Gen. Genet. 248, 735–743 (1995). https://doi.org/10.1007/BF02191714

Download citation

Key words

  • Iron transport
  • Pseudobactin
  • Outer membrane receptor
  • Pseudomonas