1887

Abstract

The type strain, ATCC 19606, secretes acinetobactin, a catechol siderophore highly related to the iron chelator anguibactin produced by the fish pathogen (). This paper reports the initial characterization of the genes and gene products involved in the acinetobactin-mediated iron-acquisition process. Insertional mutagenesis resulted in the isolation of several derivatives whose ability to grow in medium containing the iron chelator 2,2′-dipyridyl was affected. One of the insertions disrupted a gene encoding a predicted outer-membrane protein, named BauA, highly similar to FatA, the receptor for ferric anguibactin. Immunological relatedness of BauA with FatA was confirmed by Western blot analysis. Another transposon insertion was mapped to a gene encoding a protein highly similar to FatD, the permease component of the anguibactin transport system. Further DNA sequencing and nucleotide sequence analysis revealed that these 19606 genes are part of a polycistronic locus that contains the ORFs. While the translation products of , -, - and - are highly related to the FatDCBA iron-transport proteins, the product of is related to the ATPase component of Gram-positive ATP-binding cassette (ABC) transport systems. This entire locus is flanked by genes encoding predicted proteins related to AngU and AngN, proteins required for the biosynthesis of anguibactin. These protein similarities, as well as the structural similarity of anguibactin and acinetobactin, suggested that these two siderophores could be utilized by both bacterial strains, a possibility that was confirmed by siderophore utilization bioassays. Taken together, these results demonstrate that these pathogens, which cause serious infections in unrelated hosts, express very similar siderophore-mediated iron-acquisition systems.

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2004-11-01
2019-11-14
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References

  1. Actis, L. A., Potter, S. & Crosa, J. H. ( 1985; ). Iron-regulated outer membrane protein OM2 of Vibrio anguillarum is encoded by virulence plasmid pJM1. J Bacteriol 161, 736–742.
    [Google Scholar]
  2. Actis, L. A., Fish, W., Crosa, J. H., Kellerman, K., Ellenberger, S., Hauser, F. & Sanders-Loehr, J. ( 1986; ). Characterization of anguibactin, a novel siderophore from Vibrio anguillarum 775(pJM1). J Bacteriol 167, 57–65.
    [Google Scholar]
  3. Actis, L. A., Tolmasky, M. E., Farrell, D. & Crosa, J. H. ( 1988; ). Genetic and molecular characterization of essential components of the Vibrio anguillarum plasmid-mediated iron-transport system. J Biol Chem 263, 2853–2860.
    [Google Scholar]
  4. Actis, L. A., Tolmasky, M. E., Crosa, L. M. & Crosa, J. H. ( 1995; ). Characterization and regulation of the expression of FatB, an iron transport protein encoded by the pJM1 virulence plasmid. Mol Microbiol 17, 197–204.[CrossRef]
    [Google Scholar]
  5. Actis, L. A., Smoot, J. C., Barancin, C. E. & Findlay, R. H. ( 1999; ). Comparison of differential plating media and two chromatography techniques for the detection of histamine production in bacteria. J Microbiol Methods 39, 79–90.[CrossRef]
    [Google Scholar]
  6. Anstey, N. M., Currie, B. J. & Withnall, K. M. ( 1991; ). Community-acquired Acinetobacter pneumonia in the Northern Territory of Australia. Clin Infect Dis 14, 83–91.
    [Google Scholar]
  7. Arnow, L. ( 1937; ). Colorimetric determination of the components of 3,4-dihydroxyphenylalanine-tyrosine mixtures. J Biol Chem 118, 531–537.
    [Google Scholar]
  8. Bergogne-Berenzin, E. & Towner, K. J. ( 1996; ). Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features. Clin Microbiol Rev 9, 148–165.
    [Google Scholar]
  9. Bergogne-Berenzin, E., Joly-Guillou, M. L. & Towner, K. J. ( 1996; ). History and importance of Acinetobacter spp., role in infection, treatment and cost implications. In Acinetobacter: Microbiology, Epidemiology, Infections, Management, pp. 2–12. Edited by E. Bergogne-Berenzin, M. L. Joly-Guillou & K. J. Towner. Boca Raton, FL: CRC Press.
  10. Birnboim, H. C. & Doly, J. ( 1979; ). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7, 1513–1523.[CrossRef]
    [Google Scholar]
  11. Bouvet, P. J. M. & Grimont, P. A. D. ( 1986; ). Taxonomy of the genus Acinetobacter with the recognition of Acinetobacter baumannii sp. nov., Acinetobacter haemolyticus sp. nov., Acinetobacter johnsonii sp. nov., and Acinetobacter junii sp. nov. and emended descriptions of Acinetobacter calcoaceticus and Acinetobacter lwoffii. Int J Syst Bacteriol 36, 228–240.[CrossRef]
    [Google Scholar]
  12. Bradford, M. ( 1976; ). A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal Biochem 72, 249–252.
    [Google Scholar]
  13. Butterton, J. R., Choi, M. H., Watnick, P. I., Carroll, P. A. & Calderwood, S. B. ( 2000; ). Vibrio cholerae VibF is required for vibriobactin synthesis and is a member of the family of nonribosomal peptide synthetases. J Bacteriol 182, 1731–1738.[CrossRef]
    [Google Scholar]
  14. Chenault, S. S. & Earhart, C. F. ( 1991; ). Organization of genes encoding membrane proteins of the Escherichia coli ferrienterobactin permease. Mol Microbiol 5, 1405–1413.[CrossRef]
    [Google Scholar]
  15. Crosa, J. H. ( 1980; ). A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature 284, 566–568.[CrossRef]
    [Google Scholar]
  16. Crosa, J. H. ( 1997; ). Signal transduction and transcriptional and posttranscriptional control of iron-regulated genes in bacteria. Microbiol Mol Biol Rev 61, 319–336.
    [Google Scholar]
  17. Crosa, J. H. & Walsh, C. T. ( 2002; ). Genetics and assembly line enzymology of siderophore biosynthesis in bacteria. Microbiol Mol Biol Rev 66, 223–249.[CrossRef]
    [Google Scholar]
  18. Crosa, J. H., Hodges, L. & Schiewe, M. ( 1980; ). Curing of a plasmid is correlated with an attenuation of virulence in the marine fish pathogen Vibrio anguillarum. Infect Immun 27, 897–902.
    [Google Scholar]
  19. Crosa, J. H., Tolmasky, M. E., Actis, L. A. & Falkow, S. ( 1994; ). Plasmids. In Methods for General and Molecular Bacteriology, pp. 365–386. Edited by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg. Washington, DC: American Society for Microbiology.
  20. Di Lorenzo, M., Stork, M., Tolmasky, M. E. & 9 other authors ( 2003; ). Complete sequence of virulence plasmid pJM1 from the marine fish pathogen Vibrio anguillarum strain 775. J Bacteriol 185, 5822–5830.[CrossRef]
    [Google Scholar]
  21. Dorsey, C. W., Tomaras, A. P. & Actis, L. A. ( 2002; ). Genetic and phenotypic analysis of Acinetobacter baumannii insertion derivatives generated with a transposome system. Appl Environ Microbiol 68, 6353–6360.[CrossRef]
    [Google Scholar]
  22. Dorsey, C. W., Beglin, M. S. & Actis, L. A. ( 2003a; ). Detection and analysis of iron uptake components expressed by Acinetobacter baumannii clinical isolates. J Clin Microbiol 41, 4188–4193.[CrossRef]
    [Google Scholar]
  23. Dorsey, C. W., Tolmasky, M. E., Crosa, J. H. & Actis, L. A. ( 2003b; ). Genetic organization of an Acinetobacter baumannii chromosomal region harbouring genes related to siderophore biosynthesis and transport. Microbiology 149, 1227–1238.[CrossRef]
    [Google Scholar]
  24. Echenique, J. R., Arienti, H., Tolmasky, M. E., Read, R., Staneloni, J., Crosa, J. H. & Actis, L. A. ( 1992; ). Characterization of a high-affinity iron transport system in Acinetobacter baumannii. J Bacteriol 174, 7670–7679.
    [Google Scholar]
  25. Feinberg, A. P. & Vogelstein, B. ( 1983; ). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132, 6–13.[CrossRef]
    [Google Scholar]
  26. Furrer, J. L., Sanders, D. N., Hook-Barnard, I. G. & McIntosh, M. A. ( 2002; ). Export of the siderophore enterobactin in Escherichia coli: involvement of a 43 kDa membrane exporter. Mol Microbiol 44, 1225–1234.[CrossRef]
    [Google Scholar]
  27. Graber, K., Smoot, L. M. & Actis, L. A. ( 1998; ). Expression of iron binding proteins and hemin binding activity in the dental pathogen Actinobacillus actinomycetemcomitans. FEMS Microbiol Lett 163, 135–142.[CrossRef]
    [Google Scholar]
  28. Jalal, M., Hossain, D., van der Helm, J., Sanders-Loehr, J., Actis, L. A. & Crosa, J. H. ( 1989; ). Structure of anguibactin, a unique plasmid-related bacterial siderophore from the fish pathogen Vibrio anguillarum. J Am Chem Soc 111, 292–296.[CrossRef]
    [Google Scholar]
  29. Krieg, N. R. & Holt, J. G. ( 1984; ). Bergey's Manual of Systematic Bacteriology, vol. 1. Baltimore, MD: Williams & Wilkins.
  30. Leong, S., Ditta, G. S. & Helinski, D. R. ( 1982; ). Heme biosynthesis in Rhizobium: identification of a cloned gene coding for an amino levulininc acid synthetase from Rhizobium meliloti. J Biol Chem 257, 8724–8730.
    [Google Scholar]
  31. Meade, H. M., Long, S. R., Ruvkum, S. E., Brown, S. E. & Ausubel, F. M. ( 1982; ). Physical and genetic characterization of symbiotic and auxotrophic mutants Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol 149, 114–122.
    [Google Scholar]
  32. Mihara, K., Tanabe, T., Yamakawa, Y., Funahashi, T., Nakao, N., Narimatsu, N. & Yamamoto, S. ( 2004; ). Identification and transcriptional organization of a gene cluster involved in biosynthesis and transport of acinetobactin, a siderophore produced by Acinetobacter baumannii ATCC 19606T. Microbiology 150, 2587–2597.[CrossRef]
    [Google Scholar]
  33. Miller, J. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  34. Neilands, J. ( 1981; ). Iron absorption and transport in microorganisms. Annu Rev Nutr 1, 27–46.[CrossRef]
    [Google Scholar]
  35. Nelson, K., Weinel, C., Paulsen, I. & 40 other authors ( 2002; ). Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environ Microbiol 4, 799–808.[CrossRef]
    [Google Scholar]
  36. Olmsted, J. B. ( 1981; ). Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem 256, 11955–11957.
    [Google Scholar]
  37. Sambrook, J. & Russell, D. W. ( 2001; ). Molecular Cloning. A Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  38. Schweizer, H. P. & Hoang, T. T. ( 1995; ). An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosa. Gene 158, 15–22.[CrossRef]
    [Google Scholar]
  39. Schwyn, B. & Neilands, J. B. ( 1987; ). Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160, 47–56.[CrossRef]
    [Google Scholar]
  40. Shea, J. E., Hensel, M., Gleeson, C. & Holden, D. W. ( 1996; ). Identification of a virulence locus encoding a second type III secretion system in Salmonella typhimurium. Proc Natl Acad Sci U S A 93, 2593–2597.[CrossRef]
    [Google Scholar]
  41. Tolmasky, M. E., Actis, L. A. & Crosa, J. H. ( 1988; ). Genetic analysis of the iron uptake region of the Vibrio anguillarum plasmid pJM1: molecular cloning of genetic determinants encoding a novel trans activator of siderophore biosynthesis. J Bacteriol 170, 1913–1919.
    [Google Scholar]
  42. Tolmasky, M. E., Actis, L. A. & Crosa, J. H. ( 1995; ). A histidine decarboxylase gene encoded by the Vibrio anguillarum plasmid pJM1 is essential for virulence: histamine is a precursor in the biosynthesis of anguibactin. Mol Microbiol 15, 87–95.[CrossRef]
    [Google Scholar]
  43. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76, 4350–4354.[CrossRef]
    [Google Scholar]
  44. Waldbeser, L., Tolmasky, M. E., Actis, L. A. & Crosa, J. H. ( 1993; ). Mechanisms for negative regulation by iron of the fatA outer membrane protein gene expression in Vibrio anguillarum 775. J Biol Chem 268, 10433–10439.
    [Google Scholar]
  45. Walter, M., Potter, S. & Crosa, J. H. ( 1983; ). Iron uptake system mediated by Vibrio anguillarum plasmid pJM1. J Bacteriol 156, 880–887.
    [Google Scholar]
  46. Wilchek, M. & Bayer, E. A. ( 1988; ). The avidin-biotin complex in bioanalytical applications. Anal Biochem 171, 1–32.[CrossRef]
    [Google Scholar]
  47. Wu, C.-J. & Janssen, G. R. ( 1997; ). Expression of a streptomycete leaderless mRNA encoding chloramphenicol acetyltransferase in Escherichia coli. J Bacteriol 179, 6824–6830.
    [Google Scholar]
  48. Yamamoto, S., Okujo, N. & Sakakibara, Y. ( 1994; ). Isolation and structure elucidation of acinetobactin, a novel siderophore from Acinetobacter baumannii. Arch Microbiol 162, 249–252.
    [Google Scholar]
  49. Yanisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene 33, 103–109.[CrossRef]
    [Google Scholar]
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