Skip to main content
Log in

Molecular analysis and nucleotide sequence of the envCD operon of Escherichia coli

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The chromosomal DNA insert in plasmid pJK131, which complements the phenotypic defects associated with a mutation in the envC gene of Escherichia coli strain PM61, was sequenced. The analysis of the nucleotide sequence revealed two open reading frames (ORFs) coding for the proteins EnvC (41281 daltons) and EnvD (104415 daltons). The envC gene product is synthesized as a pre-protein and, after cleavage of a signal peptide, the mature protein is incorporated into the cytoplasmic membrane. The detection of a common transcript for both ORFs indicated the existence of an envCD operon. Deletion analysis and the generation of frameshifts demonstrated that simultaneous expression of both genes is required to complement the defects in strain PM61. Overproduction of EnvC protein appears to be lethal to Escherichia coli. The envD gene, however, could be cloned and expressed at high levels under control of the tac promotor without deleterious effects on the host.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Argos P, Rao JKM, Hargrave PA (1982) Structural prediction of membrane-bound proteins. Eur J Biochem 128:565–575

    Google Scholar 

  • Bachman BJ (1990) Linkage map of Escherichia coli K-12, Edition 8. Microbiol Rev 54:130–197

    Google Scholar 

  • Beck E, Bremer E (1980) Nucleotide sequence of the gene ompA coding the outer membrane protein II of Escherichia coli K-12. Nucleic Acids Res 8:3011–3024

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031

    Google Scholar 

  • Birkenbihl RP, Vielmetter W (1989) Complete maps of IS1, IS2, IS3, IS4, IS5, IS30 and IS150 locations in Escherichia coli K12. Mol Gen Genet 220:147–153

    Google Scholar 

  • de Boer PAJ, Cook WR, Rothfield LI (1990) Bacterial cell division. Annu Rev Genet 24:249–274

    Google Scholar 

  • Broome-Smith JK, Ioannidis I, Edelman A, Spratt BG (1988) Nucleotide sequence of the penicillin-binding protein 5 and 6 genes of Escherichia coli. Nucleic Acids Res 16:1617

    Google Scholar 

  • Donachie WD, Robinson AC (1987) Cell division: Parameter values and the process. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium: Cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 1578–1593

    Google Scholar 

  • Duffand GD, Lehnhardt SK, March PE, Inouye M (1985) Structure and function of the signal peptide. Curr Top Membranes Transport 24:65–104

    Google Scholar 

  • Filip C, Fletcher G, Wulf JL, Earhart CF (1973) Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol 115:717–722

    Google Scholar 

  • Gribskov M, Dvereux J, Burgess RR (1984) The codon preference plot: Graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res 12:539–549

    Google Scholar 

  • Gustafsson P, Nordström K, Normark S (1973) Outer penetration barrier of Escherichia coli K-12: Kinetics of the uptake of gentian violet by wild type and envelope mutants. J Bacteriol 116:893–900

    Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Google Scholar 

  • Henikoff D (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359

    Google Scholar 

  • Henrich B, Monnerjahn U, Plapp R (1990) Peptidase D gene (pepD) of Escherichia coli K-12: Nucleotide sequence, transcript mapping, and comparison with other peptidase genes. J Bacteriol 172:4641–4651

    Google Scholar 

  • Ito K, Sato T, Yura T (1977) Synthesis of the membrane proteins in E. coli. Cell 11:551–559

    Google Scholar 

  • Karibian D, Pellon G, Starka J (1981) Autolysis of a division mutant of Escherichia coli. J Gen Microbiol 126:55–61

    Google Scholar 

  • Karibian D, Michel GPF, Starka J (1985) The envC phenotype and its expression in various Escherichia coli K12 strains. FEMS Microbiol Lett 27:319–324

    Google Scholar 

  • Klein JR, Henrich B, Plapp R (1990) Molecular cloning of the envC gene of Escherichia coli. Curr Microbiol 21:341–347

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Levy SB, McMurry L (1974) Detection of an inducible membrane protein associated with R factor-mediated tetracycline resistance. Biochem Biophys Res Commun 56:1060–1068

    Google Scholar 

  • Luktenhaus J (1988) Genetic analysis of bacterial cell division. Microbiol Sci 5:88–91

    Google Scholar 

  • Luktenhaus J (1990) Regulation of cell division in E. coli. Trends Genet 6:22–25

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Meselson M, Yuan R (1968) DNA restriction enzyme from Escherichia coli. Nature 217:1110–1114

    Google Scholar 

  • Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19:269–276

    Google Scholar 

  • Michel G, Starka J (1979) Phospholipase A activity with integrated phospholipid vesicles in intact cells of an envelope mutant of Escherichia coli. FEBS Lett 108:261–265

    Google Scholar 

  • Michel G, Di Savino D, Starka J (1977) Phospholipid composition and phenotypic correction of an envC mutant of Escherichia coli. J Bacteriol 129:145–150

    Google Scholar 

  • Neu HC, Happel LA (1965) The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 240:3685–3692

    Google Scholar 

  • Oliver BD (1987) Periplasm and protein secretion. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium. Cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 56–69

    Google Scholar 

  • Osborn MJ, Munson R (1974) Separation of inner and outer membranes of Gram-negative bacteria. Methods Enzymol 31:642–653

    Google Scholar 

  • Pugsley AP (1989) Protein Targeting. Academic Press, London, pp 45–111

    Google Scholar 

  • Queen C, Korn LJ (1984) A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Res 12:581–599

    Google Scholar 

  • Rodolakis A, Thomas B, Starka J (1973) Morphological mutants of Escherichia coli. Isolation and ultrastructure of a chain-forming envC mutant. J Gen Microbiol 75:409–416

    Google Scholar 

  • Rodolakis A, Casse F, Starka J (1974) Morphological mutants of Escherichia coli K12. Mapping of the envC mutation. Mol Gen Genet 130:177–181

    Google Scholar 

  • Rothfield L, Romeo D (1971) Enzyme reactions in biological membranes. In: Rothfield L (ed) Structure and function of biological membranes. Academic Press, New York, pp 251–284

    Google Scholar 

  • Russel DR, Bennet GN (1982) Construction and analysis of in vivo activity of E. coli promoter hybrids and promoter mutants that alter the −35 to −10 spacing. Gene 20:231–243

    Google Scholar 

  • Sabatini DD, Kreibich G, Morimoto T, Adesnik M (1982) Mechanism for the incorporation of proteins in membranes and organelles. J Cell Biol 92:1–22

    Google Scholar 

  • Saier MH, Werner PK, Müller M (1990) Insertion of proteins into bacterial membranes: Mechanism, characteristics, and comparisons with the eucaryotic process. Microbiol Rev 53:333–366

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Spratt BG, Boyd A, Stoker N (1980) Defective and plaque-forming lambda transducing bacteriophage carrying penicillin-binding protein cell shape genes: genetic and physical mapping and identification of gene products from the lip-dacA-pbpA-leuS region of the Escherichia coli chromosome. J Bacteriol 143:569–581

    Google Scholar 

  • Staden R (1984) Measurements of the effects that coding for a protein has on a DNA sequence and their use for finding genes. Nucleic Acids Res 12:551–567

    Google Scholar 

  • Starkova Z, Thomas P, Starka J (1978) Morphological mutants of Escherichia coli: Nature of the permeability barrier in mon and envC cells. Ann Inst Pasteur Microbiol 129:265–284

    Google Scholar 

  • Starkova Z, Bonnaveiro N, Starka J (1981) Hydrolysis of phospholipids by phospholipase C in intact cells of wild-type and envelope mutants of Escherichia coli K12. FEBS Lett 130:261–264

    Google Scholar 

  • Stoker NG, Fairweather NF, Spratt BG (1982) Versatile low-copynumber plasmid vectors for cloning in Escherichia coli. Gene 18:335–341

    Google Scholar 

  • Wadle D, Henrich B, Plapp R (1986) Effects of mutations in genes fadR, fadE, and envC of Escherichia coli on the action of the lysis gene of bacteriophage ΦX174. Curr Microbiol 14:65–69

    Google Scholar 

  • van Wezenbeek PMGF, Hulsebos TJM, Schoenmakers JGG (1980) Nucleotide sequence of the filamentous bacteriophage M13 DNA genome: Comparison with phage fd. Gene 11:129–148

    Google Scholar 

  • Wollman EL, Jacob F (1956) Conjugation and genetic recombination in Escherichia coli K-12. Cold Spring Harbor Symp Quant Biol 21:141–162

    Google Scholar 

  • Wu HC, Tokunaga M (1986) Biogenesis of lipoproteins in bacteria. Curr Top Microbiol Immunol 125:127–158

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequence of the M13mpl8 and pUC19 vectors. Gene 33:103–119

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klein, J.R., Henrich, B. & Plapp, R. Molecular analysis and nucleotide sequence of the envCD operon of Escherichia coli . Molec. Gen. Genet. 230, 230–240 (1991). https://doi.org/10.1007/BF00290673

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00290673

Key words

Navigation