Summary
It is established that wild-type cells ofYersinia pestis absorb exogenous hemin or Congo red and thus grow as pigmented colonies at 26° C on media containing these chromatophores (Pgm+). Pgm+ isolates are known to possess a siderophore-independent mechanism of iron-transport (required for growth in iron-deficient medium) which is absent in avirulent Pgm− mutants. Production of the bacteriocin pesticin and linked invasins (Pst+) is an additional defined virulence factor of yersiniae; mutation of Pgm+,Pst− organisms to pesticin-resistance (Pstr) results in concomitant conversion to Pgm−. In this study, autoradiograms of two-dimensional gels of [35S]methionine-labeled outer membranes from Pgm− mutants were compared to those of the Pgm+,Pst+ or Pgm+,Pst− parent. An apparently single predominant peptide present in these preparations (> 10% of total membrane protein) existed as a family of iron-modifiable 17.9-kDa molecules focusing down to isoelectric points of about 4.6 and up to 5.89. Expression of eight detectable Pst+-specific peptides was not significantly influenced by exogenous iron. Pgm+ yersiniae constitutively produced pigmentation-specific peptide F and five iron-repressible peptides termed IrpA to IrpE. Typical spontaneous mutation to Pgm− resulted in loss of peptide F and IrpB-E. A rare Pgm+,Pstr mutant, selected on Congo red agar containing pesticin, also lost IrpB-E but retained peptide F. This isolate, like Pgm− mutants, failed to grow in iron-deficient medium. Regardless of phenotype, all yersiniae utilized hemin, hemopexin, myoglobin, hemoglobin, and ferritin, but not transferrin or lactoferrin, as sole sources of iron.
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References
Bagg A, Neilands JB (1987) Ferric uptake regulation protein acts as a repressor, employing iron(II) as a cofactor to bind the operator of an iron transport operon inEscherichia coli. Biochemistry 26:5471–5477
Ben-Gurion R, Hertman I (1958) Bacteriocin-like material produced by Pasteurella pestis. J Gen Microbiol 19:289–297
Ben-Gurion R, Shafferman A (1981) Essential virulence determinants of differentYersinia species are carried on a common plasmid. Plasmid 5:183–187
Blose SH (1986) The mouse INH/3T3 cell line protein database developed from computer-analyzed two-dimensional gels: key protein identification by experiments and amino acid ratios. In: Dunn MJ (ed) Electrophoresis '86: Proceedings of the Fifth Meeting of the International Electrophoresis Society. VCH Verlagsgesellschaft, Weinheim, pp 552–555
Brubaker RR (1970) Mutation rate to nonpigmentation inPasteurella pestis. J Bacteriol 98:1404–1406
Brubaker RR (1972) The genusYersinia: biochemistry and genetics of virulence. Curr Top Microbiol 57:111–158
Brubaker RR, Beesley ED, Surgalla MJ (1965)Pasteurella pestis: role of pesticin I and iron in experimental plague. Science 149:422–424
Brubaker RR, Surgalla MJ (1961) Pesticins I. Pesticin-bacterium interrelationships, and environmental factors influencing activity. J Bacteriol 82:940–949
Brubaker RR, Surgally MJ (1962) Pesticins. II. Production of pesticin I and II. J Bacteriol 84:539–545
Burrows TW (1963) Virulence of Pasteurella pestis and immunity of plague. Ergeb Mikrobiol 37:59–113
Carniel E, Mazigh D, Mollaret HH (1987) Expression of ironregulated proteins inYersinia species and their relation to virulence. Infect Immun 55:277–280
Carniel E, Antoine J-C, Guiyoule A, Guiso N, Mollaret HH (1989a) Purification, location and immunological charaterization of the iron-regulated high-molecular-weight proteins of the highly pathogenic Yersiniae. Infect Immun 57:540–545
Carniel E, Mercereau-Puijalon O, Bonnefoy S (1989b) The gene coding for the 190000-dalton iron-regulated protein ofYersinia species is present only in the highly pathogenic strains. Infect Immun 57:1211–1217
Conkell MB, Yanofsky C (1971) Influence of chromosome structure on the frequency oftonB trp deletions inEscherichia coli. J Bacteriol 105:864–872
Davies JK, Reeves P (1975) Genetics of resistance to colicins inEscherichia coli K-12: cross resistance among colicins of group B. J Bacteriol 123:96–101
Devignat R (1951) Variétés de l'espécePasteurella pestis. Nouvelle hypothése. Bull WHO 4:247–263
Ferber DM, Fowler JM, Brubaker RR (1981) Mutations to tolerance and resistance to pesticin and colicins in Escherichia coli Φ. J Bacteriol 146:506–511
Garrels JI (1979) Two-dimensional gel electrophoresis and computer analysis of proteins synthesized by clonal cell lines. J Biol Chem 254:7961–7977
Garrels JI (1983) Quantitative two-dimensional gel electrophoresis of proteins. Methods Enzymol 100:411–423
Garrels JI, Farrar JT, Burwell CB (1984) The QUEST system for computer-analyzed two-dimensional electrophoresis of proteins. In: Celis JE, Bravo R (eds) Two-dimensional gel electrophoresis or proteins. Academic Press, New York, pp 37–91
Goguen JD, Yother J, Straley SC (1984) Genetic analysis of the low calcium response inYersinia pestis Mu dl (Ap lac) insertion mutants. J Bacteriol 160:842–848
Hertman I, Ben-Gurion R (1959) A study of pesticin biosynthesis. J Gen Microbiol 21:135–143
Higuchi K, Kupferberg LL, Smith JL (1959) Studies on the nutrition and physiology ofPasteurella pestis: III. Effects of calcium ions on the growth of virulent and avirulent strains ofPasteurella pestis. J Bacteriol 77:317–321
Hu PC, Brubaker RR (1974) Characterization of pesticin: separation of antibacterial activities. J Biol Chem 249:4749–4753
Hu PC, Yang GCH, Brubaker RR (1972) Specificity, induction, and absorption of pesticin. J Bacteriol 112:212–219
Jackson S, Burrows TW (1956a) The pigmentation of Pasteurella pestis on a defined medium containing haemin. Br J Exp Pathol 37:570–576
Jackson S, Burrows TW (1956b) The virulence enhancing effect of iron on non-pigmented mutants of virulent strains ofPasteurella pestis. Br J Exp Pathol 37:577–583
Lankford CE (1973) Bacterial assimulation of iron. Crit Rev Microbiol 2:273–330
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Mehigh RJ, Brubaker RR (1989) Expression of the low-calcium response inYersinia pestis. Microb Pathog 6:203–217
Neilands JB (1972) Evolution of biological iron-binding centers. Struct Bonding 11:145–170
O'Farrell PH (1975) High-resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Osborn MJ, Gander JE, Parsi E, Carson J (1972) Mechanism of assembly of the outer membrane ofSalmonella typhimurium. J Biol Chem 247:3962–3972
Perry RD, Brubaker RR (1979) Accumulation of iron by yersiniae. Infect Immun 137:1290–1298
Portnoy DA, Blank HF, Kingsbury DT, Falkow S (1983) Genetic analysis of essential plasmid determinations of pathogenicity inYersinia pestis. J Infect Dis 148:297–304
Rohlf FJ, Sokal RR (1969) Statistical tables. WH Freeman and Co, San Francisco, pp 240–244
Sample AK, Fowler JM, Brubaker RR (1987) Modulation of the low calcium response inYersinia pestis by plasmidplasmid interaction. Microb Pathog 2:443–453
Sikkema DJ, Brubaker RR (1987) Resistance to pesticin, storage of iron and invasion of HeLa cells by yersiniae. Infect Immun 55:572–578
Sodeinde OA, Gogeuen JD (1988) Genetic analysis of the 9.5kilobase virulence plasmid ofYersinia pestis. Infect Immun 56:2143–2148
Sokal RR, Rohlf FJ (1969) Biometry: the principles and practice of statistics in biological research. Freeman WH and Co, San Francisco, pp 391–395
Spurr WA, Bonini CP (1973) Statistical analysis for business decisions. RD Irwin, Homewood, Ill, pp 292–298 and 704
Staggs L, Perry RD (1989) Characterization of an iron-responsivefur-like regulatory mechanism inYersinia pestis. Abstr Annu Meet Am Soc Microbiol D202
Straley SC, Brubaker RR (1981) Cytoplasmic and membrane proteins of yersiniae cultivated under conditions simulating mammalian intracellar environment. Proc Natl Acad Sci USA 78:1224–1228
Straley SC, Brubaker RR (1982) Localization in Yersinia pestis of peptides associated with virulence. Infect Immun 36:129–135
Surgalla MJ, Bessley ED (1969) Congo-red-agar plating medium for detecting pigmentation inPasteurella pestis. Appl Microbiol 18:834–837
Theil EC (1987) Ferritin: structure, gene regulation, and cellular function in animals, plants, and microorganisms. Annu Rev Biochem 56:289–315
Une T, Brubaker RR (1984) In vivo comparison of avirulent Vwa− and Pgm− or Pstr phenotypes of yersiniae. Infect Immun 43:895–900
van Asbeck BS, Verhoef J (1983) Iron and host defense. Eur J Clin Microbiol 2:6–10
Wake A, Misawa M, Matsui A (1975) Siderochrome production byYersinia pestis and its relation to virulence. Infect Immun 12:1211–1213
Waring WS, Werkman CH (1942) Growth of bacteria in an iron-free medium. Arch Biochem 1:303–310
Wee S, Neilands JB, Bittner ML, Hemming BC, Haymore BL, Seethram R (1988) Expression, isolation, and properties of Fur (ferric uptake regulation) protein ofEscherichia coli. Biol Metals 1:62–68
Weinberg ED (1974) Iron and susceptibility to infectious disease. Science 184:952–956
Wookey P (1982) ThetonB gene product inEscherichia coli. FEBS Lett 139:145–153
Zahorchak RJ, Brubaker RR (1982) Effect of exogenous nucleotides on Ca2+ dependence and V antigen synthesis inYersinia pestis. Infect Immun 38:953–959
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This is journal article no. 13025 of the Michigan Agricultural Experiment Station.
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Sikkema, D.J., Brubaker, R.R. Outer membrane peptides ofYersinia pestis mediating siderophore-independent assimilation of iron. Biol Metals 2, 174–184 (1989). https://doi.org/10.1007/BF01142557
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DOI: https://doi.org/10.1007/BF01142557