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. 1981 Sep;147(3):972–985. doi: 10.1128/jb.147.3.972-985.1981

Regulation of the OmpA outer membrane protein of Escherichia coli.

M G Beher, C A Schnaitman
PMCID: PMC216136  PMID: 7024253

Abstract

The role of lipopolysaccharide in regulating the expression of the ompA outer membrane protein gene of Escherichia coli K-12 was studied by isolating mutants defective in the biosynthesis of lipopolysaccharide and by examining transcription of lacZ in strains carrying operon fusions in which lacZ is expressed from the ompA promoter. By selecting for simultaneous resistance to phages K3 and U3, we obtained mutants defective in rfaC (biosynthesis of core heptose) and in rfaP (phosphorylation of core heptose), and both of these mutant strains failed to express OmpA protein in the outer membrane. Expression of lacZ from the ompA or by foreign ompA alleles which are not expressed in E. coli K-12. Expression was increased in strains carrying rfaC and rfaP mutations. No precursor or degraded form of OmpA protein accumulated in cells which could not express the protein in the outer membrane. This lack of accumulation of precursor was observed even in the presence of phenethyl alcohol, which caused accumulation of OmpA precursor in wild-type cells. We present a model for the regulation of this gene which is consistent with these observations and which involves modulation of transcription coupled to translation of the protein.

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Selected References

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

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Ames G. F., Spudich E. N., Nikaido H. Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. J Bacteriol. 1974 Feb;117(2):406–416. doi: 10.1128/jb.117.2.406-416.1974. [DOI] [PMC free article] [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. Bassford P. J., Jr, Diedrich D. L., Schnaitman C. L., Reeves P. Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage-resistant mutants. J Bacteriol. 1977 Aug;131(2):608–622. doi: 10.1128/jb.131.2.608-622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beher M., Pugsley A., Schnaitman C. Correlation between the expression of an Escherichia coli cell surface protein and the ability of the protein to bind to lipopolysaccharide. J Bacteriol. 1980 Jul;143(1):403–410. doi: 10.1128/jb.143.1.403-410.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benz E. W., Jr, Wydro R. M., Nadal-Ginard B., Dina D. Moloney murine sarcoma proviral DNA is a transcriptional unit. Nature. 1980 Dec 25;288(5792):665–669. doi: 10.1038/288665a0. [DOI] [PubMed] [Google Scholar]
  7. Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
  8. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  9. Chumley F. G., Menzel R., Roth J. R. Hfr formation directed by tn10. Genetics. 1979 Apr;91(4):639–655. doi: 10.1093/genetics/91.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Datta D. B., Krämer C., Henning U. Diploidy for a structural gene specifying a major protein of the outer cell envelope membrane from Escherichia coli K-12. J Bacteriol. 1976 Dec;128(3):834–841. doi: 10.1128/jb.128.3.834-841.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Diedrich D. L., Summers A. O., Schnaitman C. A. Outer membrane proteins of Escherichia coli. V. Evidence that protein 1 and bacteriophage-directed protein 2 are different polypeptides. J Bacteriol. 1977 Aug;131(2):598–607. doi: 10.1128/jb.131.2.598-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dröge W., Ruschmann E., Lüderitz O., Westphal O. Biochemical studies on lipopolysaccharides of Salmonella R mutants. 4. Phosphate groups linked to heptose units and their absence in some R lipopolysaccharides. Eur J Biochem. 1968 Mar;4(1):134–138. doi: 10.1111/j.1432-1033.1968.tb00183.x. [DOI] [PubMed] [Google Scholar]
  14. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  15. Gmeiner J., Schlecht S. Molecular organization of the outer membrane of Salmonella typhimurium. Eur J Biochem. 1979 Feb 1;93(3):609–620. doi: 10.1111/j.1432-1033.1979.tb12861.x. [DOI] [PubMed] [Google Scholar]
  16. Halegoua S., Inouye M. Translocation and assembly of outer membrance proteins of Escherichia coli. Selective accumulation of precursors and novel assembly intermediates caused by phenethyl alcohol. J Mol Biol. 1979 May 5;130(1):39–61. doi: 10.1016/0022-2836(79)90551-5. [DOI] [PubMed] [Google Scholar]
  17. Hall M. N., Silhavy T. J. Transcriptional regulation of Escherichia coli K-12 major outer membrane protein 1b. J Bacteriol. 1979 Nov;140(2):342–350. doi: 10.1128/jb.140.2.342-350.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnston H. M., Barnes W. M., Chumley F. G., Bossi L., Roth J. R. Model for regulation of the histidine operon of Salmonella. Proc Natl Acad Sci U S A. 1980 Jan;77(1):508–512. doi: 10.1073/pnas.77.1.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jousimies H., Mäkelä P. H. Genetic analysis of Salmonella minnesota R mutants with defects in the biosynthesis of the lipopolysaccharide core. J Bacteriol. 1974 Sep;119(3):753–759. doi: 10.1128/jb.119.3.753-759.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Keller E. B., Calvo J. M. Alternative secondary structures of leader RNAs and the regulation of the trp, phe, his, thr, and leu operons. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6186–6190. doi: 10.1073/pnas.76.12.6186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Koplow J., Goldfine H. Alterations in the outer membrane of the cell envelope of heptose-deficient mutants of Escherichia coli. J Bacteriol. 1974 Feb;117(2):527–543. doi: 10.1128/jb.117.2.527-543.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Lee D. R., Schnaitman C. A., Pugsley A. P. Chemical heterogeneity of major outer membrane pore proteins of Escherichia coli. J Bacteriol. 1979 Jun;138(3):861–870. doi: 10.1128/jb.138.3.861-870.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Manning P. A., Puspurs A., Reeves P. Outer membrane of Escherichia coli K-12: isolation of mutants with altered protein 3A by using host range mutants of bacteriophage K3. J Bacteriol. 1976 Sep;127(3):1080–1084. doi: 10.1128/jb.127.3.1080-1084.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Movva N. R., Nakamura K., Inouye M. Regulatory region of the gene for the ompA protein, a major outer membrane protein of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3845–3849. doi: 10.1073/pnas.77.7.3845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mühlradt P., Risse H. J., Lüderitz O., Westphal O. Biochemical studies on lipopolysaccharides of Salmonella R mutants 5. Evidence for a phosphorylating enzyme in lipopolysaccharide biosynthesis. Eur J Biochem. 1968 Apr 3;4(2):139–145. doi: 10.1111/j.1432-1033.1968.tb00184.x. [DOI] [PubMed] [Google Scholar]
  27. Nakamura K., Pirtle R. M., Pirtle I. L., Takeishi K., Inouye M. Messenger ribonucleic acid of the lipoprotein of the Escherichia coli outer membrane. II. The complete nucleotide sequence. J Biol Chem. 1980 Jan 10;255(1):210–216. [PubMed] [Google Scholar]
  28. Ornellas E. P., Stocker B. A. Relation of lipopolysaccharide character to P1 sensitivity in Salmonella typhimurium. Virology. 1974 Aug;60(2):491–502. doi: 10.1016/0042-6822(74)90343-2. [DOI] [PubMed] [Google Scholar]
  29. Oxender D. L., Zurawski G., Yanofsky C. Attenuation in the Escherichia coli tryptophan operon: role of RNA secondary structure involving the tryptophan codon region. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5524–5528. doi: 10.1073/pnas.76.11.5524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sanderson K. E., Van Wyngaarden J., Lüderitz O., Stocker B. A. Rough mutants of Salmonella typhimurium with defects in the heptose region of the lipopolysaccharide core. Can J Microbiol. 1974 Aug;20(8):1127–1134. doi: 10.1139/m74-175. [DOI] [PubMed] [Google Scholar]
  31. Sanger F., Air G. M., Barrell B. G., Brown N. L., Coulson A. R., Fiddes C. A., Hutchison C. A., Slocombe P. M., Smith M. Nucleotide sequence of bacteriophage phi X174 DNA. Nature. 1977 Feb 24;265(5596):687–695. doi: 10.1038/265687a0. [DOI] [PubMed] [Google Scholar]
  32. Schweizer M., Hindennach I., Garten W., Henning U. Major proteins of the Escherichia coli outer cell envelope membrane. Interaction of protein II with lipopolysaccharide. Eur J Biochem. 1978 Jan 2;82(1):211–217. doi: 10.1111/j.1432-1033.1978.tb12013.x. [DOI] [PubMed] [Google Scholar]
  33. Tomana M., Niedermeier W., Spivey C. Microdetermination of monosaccharides in glycoproteins by gas-liquid chromatography. Anal Biochem. 1978 Aug 15;89(1):110–118. doi: 10.1016/0003-2697(78)90731-5. [DOI] [PubMed] [Google Scholar]

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