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. 1985 Nov;164(2):585–590. doi: 10.1128/jb.164.2.585-590.1985

Domains involved in osmoregulation of the ompF gene in Escherichia coli.

K Inokuchi, M Itoh, S Mizushima
PMCID: PMC214292  PMID: 3902788

Abstract

Expression of the ompF gene, which is under the control of the OmpR protein, is regulated by the osmolarity of the medium. To study the mechanism of osmoregulation, plasmids carrying two different types of chimeric genes were constructed. In one type, the coding region of the ompF gene was linked to the trp promoter (trpPO) preceding ompF, and in the other type the ompF upstream region, mostly composed of the region for regulation by OmpR and the promoter region, was linked to the lacZ gene by protein fusion. Expression of beta-galactosidase by the lacZ chimeric gene was OmpR dependent and osmoregulated as sensitively as that of the intact ompF gene. In the ompR20 background the direction of osmoregulation was opposite that of normal osmoregulation, as was the direction of osmoregulation of the intact ompF gene. Osmoregulation was also observed with trpPO-ompF chimeric genes. However, the regulation was not as sensitive to the osmolarity of the medium as was regulation of the intact ompF gene and was independent of OmpR. These results suggest that OmpR-dependent osmoregulation played a primary role in the osmoregulation of ompF expression and that ompR-independent osmoregulation most likely did not play a crucial role. Studies with a series of trpPO-ompF chimeric genes also suggest that the untranslated leader region, about 100 base pairs in length, between the transcription initiation site and the initiation codon was not required for osmoregulation.

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  1. Alphen W. V., Lugtenberg B. Influence of osmolarity of the growth medium on the outer membrane protein pattern of Escherichia coli. J Bacteriol. 1977 Aug;131(2):623–630. doi: 10.1128/jb.131.2.623-630.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Datta D. B., Arden B., Henning U. Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors. J Bacteriol. 1977 Sep;131(3):821–829. doi: 10.1128/jb.131.3.821-829.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DiRienzo J. M., Nakamura K., Inouye M. The outer membrane proteins of Gram-negative bacteria: biosynthesis, assembly, and functions. Annu Rev Biochem. 1978;47:481–532. doi: 10.1146/annurev.bi.47.070178.002405. [DOI] [PubMed] [Google Scholar]
  5. Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Foulds J. TolF locus in Escherichia coli: chromosomal location and relationship to loci cmlB and tolD. J Bacteriol. 1976 Nov;128(2):604–608. doi: 10.1128/jb.128.2.604-608.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hall M. N., Silhavy T. J. Genetic analysis of the ompB locus in Escherichia coli K-12. J Mol Biol. 1981 Sep 5;151(1):1–15. doi: 10.1016/0022-2836(81)90218-7. [DOI] [PubMed] [Google Scholar]
  8. Hall M. N., Silhavy T. J. The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli K12. J Mol Biol. 1981 Feb 15;146(1):23–43. doi: 10.1016/0022-2836(81)90364-8. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Inokuchi K., Furukawa H., Nakamura K., Mizushima S. Characterization by deletion mutagenesis in vitro of the promoter region of ompF, a positively regulated gene of Escherichia coli. J Mol Biol. 1984 Sep 25;178(3):653–668. doi: 10.1016/0022-2836(84)90243-2. [DOI] [PubMed] [Google Scholar]
  11. Kawaji H., Mizuno T., Mizushima S. Influence of molecular size and osmolarity of sugars and dextrans on the synthesis of outer membrane proteins O-8 and O-9 of Escherichia coli K-12. J Bacteriol. 1979 Dec;140(3):843–847. doi: 10.1128/jb.140.3.843-847.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Matsuyama S., Inokuchi K., Mizushima S. Promoter exchange between ompF and ompC, genes for osmoregulated major outer membrane proteins of Escherichia coli K-12. J Bacteriol. 1984 Jun;158(3):1041–1047. doi: 10.1128/jb.158.3.1041-1047.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matsuyama S., Mizushima S. Construction and characterization of a deletion mutant lacking micF, a proposed regulatory gene for OmpF synthesis in Escherichia coli. J Bacteriol. 1985 Jun;162(3):1196–1202. doi: 10.1128/jb.162.3.1196-1202.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mizuno T., Chou M. Y., Inouye M. A comparative study on the genes for three porins of the Escherichia coli outer membrane. DNA sequence of the osmoregulated ompC gene. J Biol Chem. 1983 Jun 10;258(11):6932–6940. [PubMed] [Google Scholar]
  16. Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mizuno T., Chou M. Y., Inouye M. DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro-OmpC protein of Escherichia coli. FEBS Lett. 1983 Jan 10;151(1):159–164. doi: 10.1016/0014-5793(83)80364-0. [DOI] [PubMed] [Google Scholar]
  18. Mizuno T., Kageyama M. Separation and characterization of the outer membrane of Pseudomonas aeruginosa. J Biochem. 1978 Jul;84(1):179–191. doi: 10.1093/oxfordjournals.jbchem.a132106. [DOI] [PubMed] [Google Scholar]
  19. Mutoh N., Inokuchi K., Mizushima S. Amino acid sequence of the signal peptide of OmpF, a major outer membrane protein of Escherichia coli. FEBS Lett. 1982 Jan 25;137(2):171–174. doi: 10.1016/0014-5793(82)80341-4. [DOI] [PubMed] [Google Scholar]
  20. Mutoh N., Nagasawa T., Mizushima S. Specialized transducing bacteriophage lambda carrying the structural gene for a major outer membrane matrix protein of Escherichia coli K-12. J Bacteriol. 1981 Feb;145(2):1085–1090. doi: 10.1128/jb.145.2.1085-1090.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nakae T., Ishii J., Tokunaga M. Subunit structure of functional porin oligomers that form permeability channels in the other membrane of Escherichia coli. J Biol Chem. 1979 Mar 10;254(5):1457–1461. [PubMed] [Google Scholar]
  22. Nara F., Inokuchi K., Matsuyama S., Mizushima S. Mutation causing reverse osmoregulation of synthesis of OmpF, a major outer membrane protein of Escherichia coli. J Bacteriol. 1984 Aug;159(2):688–692. doi: 10.1128/jb.159.2.688-692.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nikaido H., Nakae T. The outer membrane of Gram-negative bacteria. Adv Microb Physiol. 1979;20:163–250. doi: 10.1016/s0065-2911(08)60208-8. [DOI] [PubMed] [Google Scholar]
  24. Nogami T., Mizushima S. Outer membrane porins are important in maintenance of the surface structure of Escherichia coli cells. J Bacteriol. 1983 Oct;156(1):402–408. doi: 10.1128/jb.156.1.402-408.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ozawa Y., Mizushima S. Regulation of outer membrane porin protein synthesis in Escherichia coli K-12: ompF regulates the expression of ompC. J Bacteriol. 1983 May;154(2):669–675. doi: 10.1128/jb.154.2.669-675.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ramakrishnan G., Ikenaka K., Inouye M. Uncoupling of osmoregulation of the Escherichia coli K-12 ompF gene from ompB-dependent transcription. J Bacteriol. 1985 Jul;163(1):82–87. doi: 10.1128/jb.163.1.82-87.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Taylor R. K., Hall M. N., Silhavy T. J. Isolation and characterization of mutations altering expression of the major outer membrane porin proteins using the local anaesthetic procaine. J Mol Biol. 1983 May 25;166(3):273–282. doi: 10.1016/s0022-2836(83)80085-0. [DOI] [PubMed] [Google Scholar]
  29. Yamada H., Kitagawa M., Kawakami M., Mizushima S. The gene coding for lipoprotein signal peptidase (lspA) and that for isoleucyl-tRNA synthetase (ileS) constitute a cotranscriptional unit in Escherichia coli. FEBS Lett. 1984 Jun 11;171(2):245–248. doi: 10.1016/0014-5793(84)80496-2. [DOI] [PubMed] [Google Scholar]
  30. Yamada H., Mizushima S. Reconstitution of an ordered structure from major outer membrane constituents and the lipoprotein-bearing peptidoglycan sacculus of Escherichia coli. J Bacteriol. 1978 Sep;135(3):1024–1031. doi: 10.1128/jb.135.3.1024-1031.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yamamoto M., Nomura M., Ohsawa H., Maruo B. Identification of a temperature-sensitive asparaginyl-transfer ribonucleic acid synthetase mutant of Escherichia coli. J Bacteriol. 1977 Oct;132(1):127–131. doi: 10.1128/jb.132.1.127-131.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yu F., Ichihara S., Mizushima S. A major outer membrane protein (O-8) of Escherichia coli K-12 exists as a trimer in sodium dodecyl sulfate solution. FEBS Lett. 1979 Apr 1;100(1):71–74. doi: 10.1016/0014-5793(79)81133-3. [DOI] [PubMed] [Google Scholar]

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