Summary
The regulatory region of the ompA gene from Escherichia coli has been characterized by biochemical and genetic approaches. Two overlapping promoters, P1 and P2, organized in that order with respect to the ompA coding sequence, were identified and it was found that ompA possesses an unusually long leader region. Both P1 and P2 were active in an in vitro transcription system although S1 mapping analysis of the ompA mRNA made in vivo showed that P2 was mainly responsible for transcription of the gene. Confirmation of this was obtained by studying down-promoter mutants of ompA cloned in pSC101. These mutants were classified into two groups, deletions and insertions. The deletions, which were caused by the IS102 insertion element found in pSC101 removed the-35 regions of both P1 and P2. However, since P2 was distally situated with respect to the IS element it was less extensively damaged and it is proposed that the residual P2 sequence is responsible for the low level of expression observed. In addition to an IS102 insertion in the promoter region four IS1 insertion mutants were characterized. These had integrated at different positions in the ompA leader region and were all incompletely polar.
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References
Adhya S, Gottesman M (1978) Control of transcription termination. Ann Rev Biochem 47:967–996
Aiba H, Adhya S, de Crombrugghe B (1981) Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
Beck E, Bremer E (1980) Nucleotide sequence of the gene ompA coding the outer membrane protein II of Escherichia coli K12. Nucleic Acid Res 8:3011–3024
Berk AJ, Sharp PA (1978) Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci USA 75:1274–1278
Bernardi A, Bernardi F (1981) Complete sequence of an IS element present in pSC101. Nucleic Acid Res 9:2905–2911
Braun V (1975) Covalent lipoprotein from the outer membrane of Escherichia coli. Biocgim. Biophys Acta 415:335–377
Bremer E, Beck E, Hindennach I, Sonntag I, Henning U (1980) Cloned structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12. Mol Gen Genet 179:13–20
Bremer E, Cole ST, Hindennach I, Henning U, Beck E, Kurz C, Schaller H (1982) Export of a protein into the outer membrane of Escherichia coli K 12. Eur J Biochem 122:223–231
Brewster JM, Morgan EA (1981) Tn9 and IS1 inserts in a ribosomal ribonucleic acid operon of Escherichia coli are incompletely polar. J Bacteriol 148, 897–903
Calos MP, Miller JH (1980) Transposable elements. Cell 20:579–596
Chai T, Foulds J (1974) Demonstration of missing outer membrane protein in tolG mutants in Escherichia coli. J Mol Biol 85:465–474
Chen R, Schmidmayr W, Krämer C, Chen-Schmeisser U, Henning U (1980) Primary structure of major outer membrane protein II* (OmpA protein) of Escherichia coli. Proc Natl Acad Sci USA 77:4592–4596
Cole ST, Chen-Schmeisser U, Hindennach I, Henning U (1982a) An apparent phage-binding region of an Escherichia coli K-12 outer membrane protein (OmpA protein). J Bacteriol in press
Cole ST, Sonntag I, Henning U (1982b) Clonign and expression in Escherichia coli K-12 of the genes for major outer membrane protein OmpA from Shigella dyserteriae, Enterobacter aerogenes and Serratia mercescens. J Bacteriol 149:145–150
Collins J (1979) Cell-free synthesis of proteins coding for mobilisation functions of ColE1 and transposition functions of Tn3. Gene 6:29–42
Das A, Court D, Gottesman M, Adhya S (1977) Polarity of insertion mutations is caused by rho-mediated termination of transcription. In: Bukhari AI, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids and episomes. Cold Spring Harbor Laboratory, New York, p 93–97
Datta DB, Arden B, Henning U (1977) Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors. J Bacteriol 131:821–829
Enderman R, Krämer C, Henning U (1978) Major outer membrane proteins of Escherichia coli K-12: evidence for protein II being a transmembrane protein. FEBS Lett 86:21–24
Galas DJ, Schmitz A (1978) DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acid Res 5:3157–3170
Gay NJ, Walker JE (1981) The ATP operon-nucleotide sequence of the promoter and the genes for the membrane proteins and the delta subunit of Escherichia coli ATP Synthase. Nucleic Acid Res 9:3919–3926
Greenblatt J (1981) Regulation of transcription termination by the N gene protein of bacteriophage lambda. Cell 24:8–9
Henning U, Sonntag I, Hindennach I (1978) Mutants (ompA) affecting a major outer membrane protein of Escherichia coli K 12. Eur J Biochem 92:491–498
Henning U, Royer HD, Teather RM, Hindennach I, Hollenberg CP (1979b) Cloning of the structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12. Proc Natl Acad Sci USA 76:4360–4364
Henning U, Schwarz H, Chen R (1979b) Radio-immunological screening method for specific membrane proteins. Anal Biochem 97:153–157
Jaurin B, Grundström T, Edlund T, Normark S (1981) The E. coli β-lactamase attenuator mediates growth rate-dependent regulation. Nature 290:221–225
Kleckner N (1981) Transposable elements in prokaryotes. Ann Rev Genet 15:341–404
Lugtenberg B, Peters R, Bernheimer H, Berendsen W (1976) Influence of cultural conditions and mutations on the composition of the outer membrane proteins of Escherichia coli. Mol Gen Genet 147:251–262
Maizels N (1973) The nucleotide sequence of the lactose messenger ribonucleic acid transcribed from the UV5 promoter mutant of Escherichia coli. Proc Natl Acad Sci USA 70:3585–3598
Manning P, Puspurs A, Reeves P (1976) Outer membrane of Escherichia coli: Isolation of mutants with altered protein 3A by using host range mutants of bacteriophage K3. J Bacteriol 127:1080–1084
Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base-specific chemical cleavages. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 65. Academic Press, New York, p 497–560
Minkley EG, Pribnow D (1973) Transcription of the early region of bacteriophage T7-Selective initiation with dinucleotides. J Mol Biol 77:255–277
Mizusawa S, Gottesman S (1982) Protein degradation in E. coli: The lon gene controls the stability of sulA protein, an inducible inhibitor of cell division. Abst Ann Meet Am Soc Microbiol H 130;134
Morita M, Oka A (1979) The structure of a transcriptional unit on colicin E1 plasmid. Eur J Biochem 97:435–443
Movva NR, Nakamura K, Inouye M (1980a) Regulatory region of the gene for the OmpA protein, a major outer membrane protein of Escherichia coli. Proc Natl Acad Sci USA 77:3845–3849
Movva NR, Nakamura K, Inouye M (1980b) Gene structure of the OmpA protein, a major surface protein of Escherichia coli required for cell-cell interaction. J Mol Biol 142:317–328
Musso RE, DiLauro R, Adhya S, de Crombrugghe B (1977) Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell 12:847–854
Nakamura K, Inouye M (1979) DNA sequence of the gene for the outer membrane lipoprotein of E. coli: an extremely AT-rich promoter. Cell 18:1109–1117
Ohtsubo H, Ohtsubo E (1978) Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci USA 75:615–619
Ohtsubo H, Zerilman M, Ohtsubo E (1980) Insertion element IS102 resides in plasmid pSC101. J Bacteriol 144:131–140
Okamoto T, Sugimoto K, Sugisaki H, Takanami M (1977) DNA regions essential for the function of a bacteriophage fd promoter. Nucleic Acid Res 4:2213–2222
Osborn MJ, Wu HCP (1980) Proteins of the outer membrane of Gram negative bacteria. Ann Rev Microbiol 34:369–422
Pratt JM, Boulnois GJ, Darby V, Orr E, Wahle E, Holland IB (1981) Identification of gene products programmed by restriction endonuclease DNA fragments using an E. coli in vitro system. Nucleic Acid Res 9:4459–4474
Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Ann Rev Genet 13:319–353
Schweizer M, Henning U (1977) Action of a major outer cell envelope membrane protein in conjugation of Escherichia coli K-12. J Bacteriol 129:1651–1652
Seeburg PH, Schaller H (1975) Mapping and characterization of promoters in bacteriophages fd, f1 and M13. J Mol Biol 92:261–277
Siebenlist U, Simpson RB, Gilbert W (1980) E. coli RNA polymerase interacts homologously with two different promoters. Cell 20:269–281
Skurray RA, Hancock REW, Reeves P (1974) Con-mutants: class of mutants in Escherichia coli K-12 lacking a major cell wall protein and defective in conjugation and adsorption of a bacteriophage. J Bacteriol 119:726–735
Smith HO, Birnstiel ML (1976) Simple method for DNA restriction site mapping. Nucleic Acid Res 3:2387–2398
Sollner-Webb B, Reeder BH (1979) The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in Xenopus laevis. Cell 18:485–499
Sonntag I, Schwarz H, Hirota Y, Henning U (1978) Cell envelope and shape of Escherichia coli: multiple mutants missing the outer membrane lipoprotein and other major outer membrane proteins. J Bacteriol 136:280–285
Starlinger P (1980) IS elements and transposons. Plasmid 3:241–259
Talkington C, Pero J (1979) Distinctive nucleotide sequences of promoters recognised by RNA polymerase containing a phagecoded “σ-like” protein. Proc Natl Acad Sci USA 76:5465–5469
Van Alphen L, Havekes L, Lugtenberg B (1977) Major outer membrane protein d of Escherichia coli K-12. Purification and in vitro activity of bacteriophage K3 and F-pilus mediated conjugation. FEBS Lett 75:285–290
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Cole, S.T., Bremer, E., Hindennach, I. et al. Characterisation of the promoters for the ompA gene which encodes a major outer membrane protein of Escherichia coli . Mol Gen Genet 188, 472–479 (1982). https://doi.org/10.1007/BF00330051
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DOI: https://doi.org/10.1007/BF00330051