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J Bacteriol. 1983 Jul; 155(1): 203–212.
PMCID: PMC217670

O-antigen conversion in Pseudomonas aeruginosa PAO1 by bacteriophage D3.

Abstract

The lysogenization of Pseudomonas aeruginosa PAO by phage D3 results in derivatives which are resistant to superinfection by phage D3c by virtue of the fact that homologous phage cannot adsorb to these cells. The serologically and morphologically unrelated phage E79 showed a markedly decreased adsorption rate to the lysogen PAO(D3). Since both of these phages are lipopolysaccharide specific, these results suggested lysogenic conversion of the phage receptor. The lipopolysaccharide was extracted from strain PAO by the hot phenol-water technique, but this procedure was ineffective with PAO(D3). We developed a technique involving cold trichloroacetic acid extraction, followed by ultracentrifugation, digestion of the high-speed pellet with proteinase K, and ultimate purification on CsCl step gradients. The lipopolysaccharide from the wild type had inactivating activity against D3 and E79, whereas that from PAO(D3) inactivated neither. Chromatographic analysis indicated that the convertant lipopolysaccharide was smooth, and quantitative chemical analyses of the two preparations showed no differences in the level of the major fatty acids, amino compounds, or neutral sugars. On the other hand, sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the side chains had a decreased migration rate through the gel matrix. The application of 1H and 13C nuclear magnetic resonance spectroscopic analysis revealed that the PAO side chain is chemically identical to that of serotype O:2a,d, containing 2,3-(1-acetyl-2-methyl-2-imidazolino-5,4)-2,3-dideoxy-D-mannuronic acid, 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid, and 2-acetamido-2,6-dideoxy-D-galactose (D-fucosamine). The molecular basis of the conversion event was (i) the introduction of an acetyl group into position 4 of the fucosamine residue and a change in the bonding between trisaccharide repeating units from alpha 1 leads to 4 to beta 1 leads to 4.

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  • Bagdian G, Lüderitz O, Staub AM. Immunochemical studies on Salmonella. XI. Chemical modification correlated with conversion of group B Salmonella by bacteriophage 27. Ann N Y Acad Sci. 1966 Jun 30;133(2):405–424. [PubMed]
  • Baker PJ, Wilson JB. Hypoferremia in mice and its application to the bioassay of endotoxin. J Bacteriol. 1965 Oct;90(4):903–910. [PMC free article] [PubMed]
  • Barksdale L, Arden SB. Persisting bacteriophage infections, lysogeny, and phage conversions. Annu Rev Microbiol. 1974;28(0):265–299. [PubMed]
  • Bartell PF, Orr TE, Reese JF, Imaeda T. Interaction of Pseudomonas bacteriophage 2 with the slime polysaccharide and lipopolysaccharide of Pseudomonas aeruginosa strain B1. J Virol. 1971 Sep;8(3):311–317. [PMC free article] [PubMed]
  • Bergan T, Midtvedt T. Epidemiological markers for pseudomonas aeruginosa. Acta Pathol Microbiol Scand B. 1975 Feb;83(1):1–9. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Buck GA, Groman NB. Identification of deoxyribonucleic acid restriction fragments of beta-converting corynebacteriophages that carry the gene for diphtheria toxin. J Bacteriol. 1981 Oct;148(1):153–162. [PMC free article] [PubMed]
  • Carey KE, Krishnapillai V. Location of prophage H90 on the chromosome of Pseudomonas aeruginosa strain PAO. Genet Res. 1974 Apr;23(2):155–164. [PubMed]
  • Carey KE, Krishnapillai V. Chromosomal location of prophage J51 in Pseudomonas aeruginosa strain PAO. Genet Res. 1975 Apr;25(2):179–187. [PubMed]
  • Castillo FJ. Partial characterization of Pseudomonas phage 2 receptor. Can J Microbiol. 1980 Aug;26(8):1015–1017. [PubMed]
  • Castillo FJ, Bartell PF. Studies on the bacteriophage 2 receptors of Pseudomonas aeruginosa. J Virol. 1974 Oct;14(4):904–909. [PMC free article] [PubMed]
  • Dimitracopoulos G, Bartell PF. Phage-related surface modifications of Pseudomonas aeruginosa: effects on the biological activity of viable cells. Infect Immun. 1979 Jan;23(1):87–93. [PMC free article] [PubMed]
  • DiRienzo JM, Deneke CF, MacLeod RA. Heterogeneity and distribution of lipopolysaccharide in the cell wall of a gram-negative marine bacterium. J Bacteriol. 1978 Oct;136(1):148–157. [PMC free article] [PubMed]
  • EGAN JB, HOLLOWAY BW. Genetic studies on lysogeny in Pseudomonas aeruginosa. Aust J Exp Biol Med Sci. 1961 Feb;39:9–17. [PubMed]
  • Galanos C, Lüderitz O, Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. [PubMed]
  • Gemski P, Jr, Koeltzow DE, Formal SB. Phage conversion of Shigella flexneri group antigens. Infect Immun. 1975 Apr;11(4):685–691. [PMC free article] [PubMed]
  • Goldman RC, Leive L. Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. Eur J Biochem. 1980;107(1):145–153. [PubMed]
  • HOLLOWAY BW, COOPER GN. Lysogenic conversion in Pseudomonas aeruginosa. J Bacteriol. 1962 Dec;84:1321–1324. [PMC free article] [PubMed]
  • HOLLOWAY BW, EGAN JB, MONK M. Lysogeny in Pseudomonas aeruginosa. Aust J Exp Biol Med Sci. 1960 Aug;38:321–329. [PubMed]
  • Holloway BW, Krishnapillai V, Morgan AF. Chromosomal genetics of Pseudomonas. Microbiol Rev. 1979 Mar;43(1):73–102. [PMC free article] [PubMed]
  • Horton D, Riley DA. 31P nuclear magnetic resonance spectroscopy of lipopolysaccharides from Pseudomonas aeruginosa. Biochim Biophys Acta. 1981 Feb 6;640(3):727–733. [PubMed]
  • Hurlbert RE, Hurlbert IM. Biological and physicochemical properties of the lipopolysaccharide of Chromatium vinosum. Infect Immun. 1977 Jun;16(3):983–994. [PMC free article] [PubMed]
  • Jann B, Reske K, Jann K. Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Eur J Biochem. 1975 Dec 1;60(1):239–246. [PubMed]
  • Jarrell K, Kropinski AM. Identification of the cell wall receptor for bacteriophage E79 in Pseudomonas aeruginosa strain PAO. J Virol. 1977 Sep;23(3):461–466. [PMC free article] [PubMed]
  • Jarrell K, Kropinski AM. The chemical composition of the lipopolysaccharide from Pseudomonas aeruginosa strain PAO and a spontaneously derived rough mutant. Microbios. 1977;19(76):103–116. [PubMed]
  • Jarrell KF, Kropinski AM. Isolation and characterization of a bacteriophage specific for the lipopolysaccharide of rough derivatives of Pseudomonas aeruginosa strain PAO. J Virol. 1981 May;38(2):529–538. [PMC free article] [PubMed]
  • Jarrell KF, Kropinski AM. Pseudomonas aeruginosa bacteriophage phi PLS27-lipopolysaccharide interactions. J Virol. 1981 Nov;40(2):411–420. [PMC free article] [PubMed]
  • Key BA, Gray GW, Wilkinson SG. The purification and chemical composition of the lipopolysaccharide of Pseudomonas alcaligenes. Biochem J. 1970 Dec;120(3):559–566. [PMC free article] [PubMed]
  • Krishnapillai V, Carey KE. Chromosomal location of a prophage in Pseudomonas aeruginosa strain PAO. Genet Res. 1972 Aug;20(1):137–140. [PubMed]
  • Kropinski AM, Chan LC, Milazzo FH. The extraction and analysis of lipopolysaccharides from Pseudomonas aeruginosa strain PAO, and three rough mutants. Can J Microbiol. 1979 Mar;25(3):390–398. [PubMed]
  • Kropinski AM, Kuzio J, Angus BL, Hancock RE. Chemical and chromatographic analysis of lipopolysaccharide from an antibiotic-supersusceptible mutant of Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1982 Feb;21(2):310–319. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lányi B, Lantos J. Antigenic changes in Pseudomonas aeruginosa in vivo and after lysogenization in vitro. Acta Microbiol Acad Sci Hung. 1976;23(4):337–351. [PubMed]
  • Lehnhardt WF, Winzler RJ. Determination of neutral sugars in glycoproteins by gas-liquid chromatography. J Chromatogr. 1968 May 7;34(4):471–479. [PubMed]
  • Lindberg AA. Studies of a receptor for felix O-1 phage in Salmonella minnesota. J Gen Microbiol. 1967 Aug;48(2):225–233. [PubMed]
  • Lindberg AA, Svensson S. Salmonella typhimurium mutations conferring resistance to Felix O phage without loss of smooth character: phage attachment and immunochemical and structural analyses of lipopolysaccharides. J Gen Microbiol. 1975 Mar;87(1):11–19. [PubMed]
  • Lindberg AA, Wollin R, Gemski P, Wohlhieter JA. Interaction between bacteriophage Sf6 and Shigella flexner. J Virol. 1978 Jul;27(1):38–44. [PMC free article] [PubMed]
  • Losick R, Robbins PW. Mechanism of epsilon-15 conversion studies with a bacterial mutant. J Mol Biol. 1967 Dec 28;30(3):445–455. [PubMed]
  • Liu PV. Changes in somatic antigens of Pseudomonas aeruginosa induced by bacteriophages. J Infect Dis. 1969 Mar;119(3):237–246. [PubMed]
  • Madhubala, Prakash K, Sharma KB. Change in the "O' serotype of Pseudomonas aeruginosa after lysogenisation with bacteriophages. Indian J Med Res. 1981 May;73:686–691. [PubMed]
  • Martinez-Molina E, Olivares J. Antibiotic production by Pseudomonas reptilivora as a phage conversion. Can J Microbiol. 1979 Sep;25(9):1108–1110. [PubMed]
  • Miller RV, Ku CM. Characterization of Pseudomonas aeruginosa mutants deficient in the establishment of lysogeny. J Bacteriol. 1978 Jun;134(3):875–883. [PMC free article] [PubMed]
  • Miller RV, Pemberton JM, Richards KE. F116, D3 and G101: temperate bacteriophages of Pseudomonas aeruginosa. Virology. 1974 Jun;59(2):566–569. [PubMed]
  • Morrison DC, Leive L. Fractions of lipopolysaccharide from Escherichia coli O111:B4 prepared by two extraction procedures. J Biol Chem. 1975 Apr 25;250(8):2911–2919. [PubMed]
  • Mühlradt PF, Wray V, Lehmann V. A 31P-nuclear-magnetic-resonance study of the phosphate groups in lipopolysaccharide and lipid A from Salmonella. Eur J Biochem. 1977 Nov 15;81(1):193–203. [PubMed]
  • Munford RS, Hall CL, Rick PD. Size heterogeneity of Salmonella typhimurium lipopolysaccharides in outer membranes and culture supernatant membrane fragments. J Bacteriol. 1980 Nov;144(2):630–640. [PMC free article] [PubMed]
  • Nygren H, Dahlén G, Nilsson LA. Human complement activation by lipopolysaccharides from bacteroides oralis, fusobacterium nucleatum, and veillonella parvula. Infect Immun. 1979 Nov;26(2):391–396. [PMC free article] [PubMed]
  • Porter WH. Application of nitrous acid deamination of hexosamines to the simultaneous GLC determination of neutral and amino sugars in glycoproteins. Anal Biochem. 1975 Jan;63(1):27–43. [PubMed]
  • Rosner MR, Khorana HG, Satterthwait AC. The structure of lipopolysaccharide from a heptose-less mutant of Escherichia coli K-12. II. The application of 31P NMR spectroscopy. J Biol Chem. 1979 Jul 10;254(13):5818–5825. [PubMed]
  • Royle PL, Matsumoto H, Holloway BW. Genetic circularity of the Pseudomonas aeruginosa PAO chromosome. J Bacteriol. 1981 Jan;145(1):145–155. [PMC free article] [PubMed]
  • Schnaitman C, Smith D, de Salsas MF. Temperate Bacteriophage Which Causes the Production of a New Major Outer Membrane Protein by Escherichia coli. J Virol. 1975 May;15(5):1121–1130. [PMC free article] [PubMed]
  • Scott JE. Letter: Phosphotungstic acid. "Schiff-reactive" but not a "glycol reagent". J Histochem Cytochem. 1973 Dec;21(12):1084–1085. [PubMed]
  • SLAYTER HS, HOLLOWAY BW, HALL CE. THE STRUCTURE OF PSEUDOMONAS AERUGINOSA PHAGES B3, E79, AND F116. J Ultrastruct Res. 1964 Oct;11:274–281. [PubMed]
  • Sugiyama H. Clostridium botulinum neurotoxin. Microbiol Rev. 1980 Sep;44(3):419–448. [PMC free article] [PubMed]
  • Wright A, Kanegasaki S. Molecular aspects of lipopolysaccharides. Physiol Rev. 1971 Oct;51(4):748–784. [PubMed]

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