Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Aug;159(2):704–712. doi: 10.1128/jb.159.2.704-712.1984

New Salmonella typhimurium mutants with altered outer membrane permeability.

S Sukupolvi, M Vaara, I M Helander, P Viljanen, P H Mäkelä
PMCID: PMC215702  PMID: 6378889

Abstract

We describe three new classes of Salmonella typhimurium mutants with increased sensitivity to hydrophobic agents. In contrast to many previously described mutants, the phage sensitivity pattern of these mutants did not give any indication of defective lipopolysaccharide. Furthermore, they had no detectable changes in their phospholipid or outer membrane protein composition, and their growth rate and cell morphology were normal. Class B mutants were nearly as sensitive to novobiocin, fusidic acid, erythromycin, rifampin, and clindamycin as are deep rough (heptoseless) mutants; in addition they were sensitive to methicillin, penicillin (to which heptoseless mutants are resistant), gentian violet, and anionic and cationic detergents. Class A and C mutants had less sensitive, but characteristic phenotypes. None of the three classes were sensitive to serum bactericidal action. The class B mutation mapped between map positions 7 and 11 on the S. typhimurium chromosome, and the class C mutation mapped between positions 5 and 7. The map position for the class A mutation remained undefined, but it was separate from the class B and C mutations and, like those, did not correspond to any gene loci known to participate in the synthesis of major outer membrane constituents.

Full text

PDF
708

Images in this article

Selected References

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

  1. Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coleman W. G., Jr, Leive L. Two mutations which affect the barrier function of the Escherichia coli K-12 outer membrane. J Bacteriol. 1979 Sep;139(3):899–910. doi: 10.1128/jb.139.3.899-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies J. K., Reeves P. Genetics of resistance to colicins in Escherichia coli K-12: cross-resistance among colicins of group A. J Bacteriol. 1975 Jul;123(1):102–117. doi: 10.1128/jb.123.1.102-117.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  7. Graham A. C., Stocker B. A. Genetics of sensitivity of Salmonella species to colicin M and bacteriophages T5, T1, and ES18. J Bacteriol. 1977 Jun;130(3):1214–1223. doi: 10.1128/jb.130.3.1214-1223.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grundström T., Normark S., Magnusson K. E. Overproduction of outer membrane protein suppresses envA-induced hyperpermeability. J Bacteriol. 1980 Dec;144(3):884–890. doi: 10.1128/jb.144.3.884-890.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hancock R. E., Reeves P. Bacteriophage resistance in Escherichia coli K-12: general pattern of resistance. J Bacteriol. 1975 Mar;121(3):983–993. doi: 10.1128/jb.121.3.983-993.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hedges R. W., Datta N., Kontomichalou P., Smith J. T. Molecular specificities of R factor-determined beta-lactamases: correlation with plasmid compatibility. J Bacteriol. 1974 Jan;117(1):56–62. doi: 10.1128/jb.117.1.56-62.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Imae Y. Mitomycin C-sensitive mutant of Eschericha coli K-12. J Bacteriol. 1968 Mar;95(3):1191–1192. doi: 10.1128/jb.95.3.1191-1192.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johansson V., Aarti A., Nurminen M., Mäkelä P. H. Outer membrane protein-specific bacteriophages of Salmonella typhimurium. J Gen Microbiol. 1978 Jul;107(1):183–187. doi: 10.1099/00221287-107-1-183. [DOI] [PubMed] [Google Scholar]
  13. Kamio Y., Nikaido H. Outer membrane of Salmonella typhimurium: accessibility of phospholipid head groups to phospholipase c and cyanogen bromide activated dextran in the external medium. Biochemistry. 1976 Jun 15;15(12):2561–2570. doi: 10.1021/bi00657a012. [DOI] [PubMed] [Google Scholar]
  14. Kuo T. T., Stocker B. A. Mapping of rfa Genes in Salmonella typhimurium by ES18 and P22 Transduction and by Conjugation. J Bacteriol. 1972 Oct;112(1):48–57. doi: 10.1128/jb.112.1.48-57.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Morona R., Reeves P. The tolC locus of Escherichia coli affects the expression of three major outer membrane proteins. J Bacteriol. 1982 Jun;150(3):1016–1023. doi: 10.1128/jb.150.3.1016-1023.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nakamura H., Suganuma A. Membrane mutation associated with sensitivity to acriflavine in Escherichia coli. J Bacteriol. 1972 Apr;110(1):329–335. doi: 10.1128/jb.110.1.329-335.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nelson B. W., Roantree R. J. Analyses of lipopolysaccharides extracted from penicillin-resistant, serum-sensitive salmonella mutants. J Gen Microbiol. 1967 Aug;48(2):179–188. doi: 10.1099/00221287-48-2-179. [DOI] [PubMed] [Google Scholar]
  19. Nikaido H. Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta. 1976 Apr 16;433(1):118–132. doi: 10.1016/0005-2736(76)90182-6. [DOI] [PubMed] [Google Scholar]
  20. Nikaido H., Song S. A., Shaltiel L., Nurminen M. Outer membrane of Salmonella XIV. Reduced transmembrane diffusion rates in porin-deficient mutants. Biochem Biophys Res Commun. 1976 May 23;76(2):324–330. doi: 10.1016/0006-291x(77)90728-8. [DOI] [PubMed] [Google Scholar]
  21. Nishijima M., Raetz C. R. Membrane lipid biogenesis in Escherichia coli: identification of genetic loci for phosphatidylglycerophosphate synthetase and construction of mutants lacking phosphatidylglycerol. J Biol Chem. 1979 Aug 25;254(16):7837–7844. [PubMed] [Google Scholar]
  22. Normark S. Mutation in Escherichia coli K-12 mediating spherelike envelopes and changes tolerance to ultraviolet irradiation and some antibiotics. J Bacteriol. 1969 Jun;98(3):1274–1277. doi: 10.1128/jb.98.3.1274-1277.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nurminen M., Lounatmaa K., Sarvas M., Mäkelä P. H., Nakae T. Bacteriophage-resistant mutants of Salmonella typhimurium deficient in two major outer membrane proteins. J Bacteriol. 1976 Aug;127(2):941–955. doi: 10.1128/jb.127.2.941-955.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Palva E. T., Liljeström P., Harayama S. Cosmid cloning and transposon mutagenesis in Salmonella typhimurium using phage lambda vehicles. Mol Gen Genet. 1981;181(2):153–157. doi: 10.1007/BF00268420. [DOI] [PubMed] [Google Scholar]
  25. Palva E. T., Mäkelä P. H. Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Eur J Biochem. 1980;107(1):137–143. doi: 10.1111/j.1432-1033.1980.tb04634.x. [DOI] [PubMed] [Google Scholar]
  26. Roantree R. J., Kuo T. T., MacPhee D. G. The effect of defined lipopolysaccharide core defects upon antibiotic resistances of Salmonella typhimurium. J Gen Microbiol. 1977 Dec;103(2):223–234. doi: 10.1099/00221287-103-2-223. [DOI] [PubMed] [Google Scholar]
  27. Rowley D. Sensitivity of rough gram-negative bacteria to the bactericidal action of serum. J Bacteriol. 1968 May;95(5):1647–1650. doi: 10.1128/jb.95.5.1647-1650.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sabet S. F., Schnaitman C. A. Purification and properties of the colicin E3 receptor of Escherichia coli. J Biol Chem. 1973 Mar 10;248(5):1797–1806. [PubMed] [Google Scholar]
  29. Sanderson K. E., Ross H., Ziegler L., Mäkelä P. H. F + , Hfr, and F' strains of Salmonella typhimurium and Salmonella abony. Bacteriol Rev. 1972 Dec;36(4):608–637. doi: 10.1128/br.36.4.608-637.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, Edition VI. Microbiol Rev. 1983 Sep;47(3):410–453. doi: 10.1128/mr.47.3.410-453.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sargent M. G. Rapid fixed-time assay for penicillinase. J Bacteriol. 1968 Apr;95(4):1493–1494. doi: 10.1128/jb.95.4.1493-1494.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schlecht S., Westphal O. Untersuchungen zur Typisierung von Salmonella-R-Formen. 4. Typisierung von S. minnesota-R-Mutanten mittels Antibiotica. Zentralbl Bakteriol Orig. 1970 Apr;213(3):356–380. [PubMed] [Google Scholar]
  33. Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
  34. Schweizer M., Schwarz H., Sonntag I., Henning U. Mutational change of membrane architecture. Mutants of Escherichia coli K12 missing major proteins of the outer cell envelope membrane. Biochim Biophys Acta. 1976 Oct 19;448(3):474–491. doi: 10.1016/0005-2736(76)90301-1. [DOI] [PubMed] [Google Scholar]
  35. Smit J., Kamio Y., Nikaido H. Outer membrane of Salmonella typhimurium: chemical analysis and freeze-fracture studies with lipopolysaccharide mutants. J Bacteriol. 1975 Nov;124(2):942–958. doi: 10.1128/jb.124.2.942-958.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stocker B. A., Nurminen M., Mäkelä P. H. Mutants defective in the 33K outer membrane protein of Salmonella typhimurium. J Bacteriol. 1979 Aug;139(2):376–383. doi: 10.1128/jb.139.2.376-383.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tamaki S., Sato T., Matsuhashi M. Role of lipopolysaccharides in antibiotic resistance and bacteriophage adsorption of Escherichia coli K-12. J Bacteriol. 1971 Mar;105(3):968–975. doi: 10.1128/jb.105.3.968-975.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vaara M., Vaara T. Sensitization of Gram-negative bacteria to antibiotics and complement by a nontoxic oligopeptide. Nature. 1983 Jun 9;303(5917):526–528. doi: 10.1038/303526a0. [DOI] [PubMed] [Google Scholar]
  39. Weigand R. A., Rothfield L. I. Genetic and physiological classification of periplasmic-leaky mutants of Salmonella typhimurium. J Bacteriol. 1976 Jan;125(1):340–345. doi: 10.1128/jb.125.1.340-345.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Weigand R. A., Vinci K. D., Rothfield L. I. Morphogenesis of the bacterial division septum: a new class of septation-defective mutants. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1882–1886. doi: 10.1073/pnas.73.6.1882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilkinson R. G., Gemski P., Jr, Stocker B. A. Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J Gen Microbiol. 1972 May;70(3):527–554. doi: 10.1099/00221287-70-3-527. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES