Skip to content
1887

Microbiology Society logo Microbiology

Volume 129, Issue 11

Dye Sensitivity Correlated with Envelope Protein Changes in () Mutants of K12 Defective in the Expression of the Sex Factor F Free

Abstract

Mutations of the gene on the chromosome result in sensitivity to the dye toluidine blue and, in male cells, cause loss of F pili, producing sterility in conjugation. Compared with its parent, a strain deleted for () showed an altered sensitivity to a wide range of dyes and antibiotics which affect different intracellular processes, and hence it appeared likely that the barrier properties of the cell envelope were impaired. Unlike mutants known to be defective in LPS structure, there appeared to be no correlation between the hydrophobicity of the compounds and the sensitivity of the strain. Moreover there was no difference between and strains in their sensitivity to LPS-specific phages, and chemical and GLC analysis of LPS components revealed no difference between the two strains. Examination of outer and inner membrane proteins from isogenic strains having the deletion and the gene cloned into the plasmid pACYC184, with or without insertional inactivation of by the transposon γδ, was performed by SDS-PAGE. This revealed a number of differences in the profile of proteins from both inner and outer membranes, correlated with mutation in the gene. The gene appears to be identical to the gene, which has been shown to be required for efficient transcription of the sex factor F. It is therefore proposed that the gene product may also control the expression of chromosomal genes coding for envelope proteins.

  • Published Online:
© Society for General Microbiology 1983
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-11-3363
1983-11-01
2025-01-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/11/mic-129-11-3363.html?itemId=/content/journal/micro/10.1099/00221287-129-11-3363&mimeType=html&fmt=ahah

References

  1. Ames B. N. 1966; Assay of inorganic phosphate, total phosphate, and phosphatases. Methods in Enzymology115–118
     [Google Scholar]
  2. Beutin L., Achtman M. 1979; Two Escherichia coli chromosomal cistrons, sfrA and sfrB, which are needed for expression of F factor tra functions. Journal of Bacteriology 139:730–737
     [Google Scholar]
  3. Beutin L., Manning P.A., Achtman M., Willetts N. 1981; sfrA and sfrB products of Escherichia coli K-12 are transcriptional control factors. Journal of Bacteriology 145:840–844
     [Google Scholar]
  4. Boman H.G., Monner D.A. 1975; Characterization of lipopolysaccharides from Escherichia coliK-12 mutants. Journal of Bacteriology 121:455–464
     [Google Scholar]
  5. Boyd A., Holland I.B. 1979; Regulation of the synthesis of surface protein in the cell cycle of E. coliB/r. Cell 18:287–296
     [Google Scholar]
  6. Buxton R.S., Drury L.S. 1983; Cloning and insertional inactivation of the dye {sfrA) gene, mutation of which affects sex factor F expression and dye sensitivity of Escherichia coli K-12. Journal of Bacteriology 154:1309–1314
     [Google Scholar]
  7. Buxton R.S., Holland I.B. 1973; Genetic studies of tolerance to colicin E2 in Escherichia coliK-12.1. Re-location and dominance relationships of cet mutations. Molecular and General Genetics 127:69–88
     [Google Scholar]
  8. Buxton R.S., Hammer-Jespersen K., Hansen T.D. 1966; Insertion of bacteriophage lambda into the deo operon of Escherichia coli K-12 and isolation of plaque-forming λ deo+ transducing bacteriophages. Journal of Bacteriology 136:668–681
     [Google Scholar]
  9. Chang A.C.Y., Cohen S.N. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. Journal of Bacteriology 134:1141–1156
     [Google Scholar]
  10. Churchward G.G., Holland I.B. 1976; Envelope synthesis during the cell cycle in Escherichia coli B/r. Journal of Molecular Biology 105:245–261
     [Google Scholar]
  11. Coleman W.G., Leive L. 1979; Two mutations which affect the barrier function of the Escherichia coli K-12 outer membrane. Journal of Bacteriology 139:899–910
     [Google Scholar]
  12. Filip C., Fletcher G., Wulff J.L., Earhart C.F. 1973; Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. Journal of Bacteriology 115:717–722
     [Google Scholar]
  13. Gaffney D., Skurray R., Willetts N. 1983; Regulation of the F conjugation genes studied by hybridization and tra-lacZ fusion. Journal of Molecular Biology 168:103–122
     [Google Scholar]
  14. Galanos C., Lüderitz O., Westphal O. 1969; A new method for the extraction of R lipopolysaccharides. European Journal of Biochemistry 9:245–249
     [Google Scholar]
  15. Holme T., Lindberg A.A., Garegg P.J., Onn T. 1968; Chemical composition of cell-wall polysaccharide of rough mutants of Salmonella typhimurium. Journal of General Microbiology 52:45–54
     [Google Scholar]
  16. Kalckar H.M., Laursen P., Rapin A.M.C. 1966; Inactivation of phage C21 by various preparations from lipopolysaccharide of E. coliK-12. Proceedings of the National Academy of Sciences of the United States of America, 56:1852–1858
     [Google Scholar]
  17. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  18. Levvy G.A., Mcallan A. 1959; The N-acetylation and estimation of hexosamines. Biochemical Journal 73:127–132
     [Google Scholar]
  19. Monner D.A., Jonsson S., Boman H.G. 1971; Ampicillin-resistant mutants of Escherichia coli K-12 with lipopolysaccharide alterations affecting mating ability and susceptibility to sex-specific bacteriophages. Journal of Bacteriology 107:420–432
     [Google Scholar]
  20. Nikaido H. 1976; Outer membranes of Salmonella typhimurium.Transmembrane diffusion of some hydrophobic substances. Biochimica et biophysica acta 433:118–132
     [Google Scholar]
  21. Nikaido H. 1979; Nonsepecific transport through the outer membrane. In Bacterial Outer Membranes. Biogenesis and Functions pp. 361–407 Edited by Inouye M. New York: Wiley;
     [Google Scholar]
  22. Osborn M.J. 1963; Studies on the Gram-negative cell wall. I. Evidence for the role of 2-keto-3-deoxyoctonate in the lipopolysaccharide of Salmonella typhimurium. Proceedings of the National Academy of Sciences of the United States of America 50:499–506
     [Google Scholar]
  23. Osborn M.J., Gander J.E., Parisi E., Carson J. 1972; Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. Journal of Biological Chemistry 247:3962–3972
     [Google Scholar]
  24. Rapin A.M.C., Kalckar H.M., Alberico L. 1968; The metabolic basis for masking of receptorsites on E. coli K-12 for C21, a lipopolysaccharide core-specific phage. Archives of Biochemistry and Biophysics 128:95–105
     [Google Scholar]
  25. Roeder W., Somerville R.L. 1979; Cloning the trpR gene. Molecular and General Genetics 176:361–368
     [Google Scholar]
  26. Sanderson K.E., Stocker B.A.D. 1981; Gene rfaH, which affects lipopolysaccharide core structure in Salmonella typhimurium, is required also for expression of F-factor functions. Journal of Bacteriology 146:535–541
     [Google Scholar]
  27. Sanderson K.E., Van Wyngaarden J., Lüderitz O., Stocker B.A.D. 1974; Rough mutants of Salmonella typhimurium with defects on the hepatose region of the lipopolysaccharide core. Canadian Journal of Microbiology 20:1127–1134
     [Google Scholar]
  28. Schmidt G., Jann B., Jann K. 1969; Immunochemistry of R lipopolysaccharides of Escherichia coli. Different core regions in the lipopolysaccharides of O group 8. European Journal of Biochemistry 10:501–510
     [Google Scholar]
  29. Schmidt G., Fromme I., Mayer H. 1970a; Immunochemical studies on core lipopolysaccharides of Enterobacteriaceae of different genera. European Journal of Biochemistry 14:357–366
     [Google Scholar]
  30. Schmidt G., Jann B., Jann K. 1969b; Immuno-chemistry of R lipopolysaccharides of Escherichia coli. Studies on R mutants with an incomplete core, derived from E. coli 08:K27. European Journal of Biochemistry 16:382–392
     [Google Scholar]
  31. Stocker B.A.D., Males B.M., Takano W. 1980; Salmonella typhimurium mutants of RfaH− phenotype: genetics and antibiotic sensitivities. Journal of General Microbiology 116:17–24
     [Google Scholar]
  32. Watson G., Paigen K. 1971; Isolation and characterization of an Escherichia coli bacteriophage requiring cell wall galactose. Journal of Virology 8:669–674
     [Google Scholar]
  33. Weissbach A., Hurwitz J. 1959; The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. Journal of Biological Chemistry 234:705–709
     [Google Scholar]
/content/journal/micro/10.1099/00221287-129-11-3363
Loading
Dye Sensitivity Correlated with Envelope Protein Changes in dye (sfrA) Mutants of Escherichia coli K12 Defective in the Expression of the Sex Factor F
Microbiology 129, 3363 (1983); https://doi.org/10.1099/00221287-129-11-3363
/content/journal/micro/10.1099/00221287-129-11-3363
/content/journal/micro/10.1099/00221287-129-11-3363
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error