Skip to main content
SpringerLink
Log in

Sequence of the mglB gene from Escherichia coli K12: Comparison of wild-type and mutant galactose chemoreceptors

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

Summary

The mglB gene of Escherichia coli codes for a galactose-binding protein (GBP) that serves both as the galactose chemoreceptor and as the recognition component of the β-methylgalactoside transport system. The mglB551 mutation eliminates the chemotactic function of GBP without altering its transport or substrate-binding properties. To investigate the interaction between GBP and Trg, the chemotactic signal transducer for galactose, we sequenced the mglB genes from wild-type and mglB551 mutant strains. The mutation causes the replacement of Gly74 of GBP by Asp. This residue is located in alpha-Helix III at the tip of the P domain in the GBP tertiary structure farthest removed from the substrate-binding cleft between the P and Q domains. We conclude that Helix III must be part of, or at least adjacent to, the recognition site for Trg. Our sequence also included part of the mglA gene, which is immediately distal to mglB. The amino acid sequence deduced for the beginning of the MglA protein showed homology with a family of polypeptides that contain an ATP-binding site and are components of binding-protein-dependent transport systems.

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

Access this article

Log in via an institution

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

  • Ames GL (1986) Bacterial periplasmic transport systems: Structure, mechanism, and evolution. Annu Rev Biochem 55:397–425

    Google Scholar 

  • Anraku Y (1967) The reduction and restoration of galactose transport in osmotically shocked cells of Escherichia coli. J Biol Chem 242:793–800

    Google Scholar 

  • Argos P, Mahoney WC, Hermondson MA, Hanei M (1981) Structural predictions of sugar binding proteins functional in chemotaxis and transport. J Biol Chem 256:4357–4361

    Google Scholar 

  • Bavoil P, Hoffnung M, Nikaido H (1980) Identification of a cytoplasmic membrane-associated component of the maltose transport system of E. coli. J Biol Chem 255:8366–8369

    Google Scholar 

  • Bolivar F, Rodriguez RL, Green PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Boos W (1969) The galactose binding protein and its relationship to the β-methylgalactoside permease from Escherichia coli. Eur J Biochem 10:66–73

    Google Scholar 

  • Boos W (1972) Transport properties of the galactose-binding protein of Escherichia coli: Substrate-induced conformational change. J Biol Chem 247:917–924

    Google Scholar 

  • Boos W, Steinacher I, Engelhardt-Altendorf D (1981) Mapping of mglB, the structural gene of the galactose-binding protein of Escherichia coli. Mol Gen Genet 184:508–518

    Google Scholar 

  • Brass JM, Manson MD (1984) Reconstitution of maltose chemotaxis in Escherichia coli by addition of maltose binding-protein to calcium-treated cells of maltose regulon mutants. J Bacteriol 157:93–99

    Google Scholar 

  • Brass JM, Ehmann U, Bukau B (1983) Reconstitution of maltose transport in Escherichia coli: conditions effecting import of maltose-binding protein into the periplasm of calcium treated cells. J Bacteriol 155:97–106

    Google Scholar 

  • Bremer E, Silhavy TJ, Weisemann JM, Weinstock GM (1984) λplacMu: a transposable derivative of phage λ for creating lacZ protein fusions in a single step. J Bacteriol 158:1084–1093

    Google Scholar 

  • Casadaban M (1976) Transposition and fusion of the lac genes to selected promotors in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol 104:541–555

    Google Scholar 

  • Dahl M, Manson MD (1985) Interspecific reconstitution of maltose transport and chemotaxis in Escherichia coli with maltose-binding protein from various enteric bacteria. J Bacteriol 164:1057–1063

    Google Scholar 

  • Duplay P, Bedouelle H, Fowler A, Zabin I, Saurin W, Hofnung M (1984) Sequences of the malE gene and its product, the maltose-binding protein of Escherichia coli K12. J Biol Chem 259:10606–10613

    Google Scholar 

  • Ebright RH, Cossart P, Gicquel-Sanzey B, Beckwith J (1984) Mutations that alter the DNA sequence specificity of the catabolite gene activator protein of E. coli. Nature 311:232–235

    Google Scholar 

  • Engelhardt-Altendorf D (1983) Mechanismus der Signalübertragung während der Chemotaxis bei Escherichia coli. Doctoral thesis, University of Konstanz, FRG

  • Gilliland GL, Quiocho FA (1981) Structure of the L-arabinose-binding protein from Escherichia coli at 2.4 Å resolution. J Mol Biol 146:341–362

    Google Scholar 

  • Grosjean H, Fiers W (1982) Preferential codon usage in prokaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene 18:199–209

    Google Scholar 

  • Hall MN, Gabay J, Débarbouillé, Schwartz M (1982) A role for secondary structure in the control of translation initiation. Nature 295:616–618

    Google Scholar 

  • Harayama S, Bollinger J, Iino T, Hazelbauer GL (1983) Characterization of the mgl operon of Escherichia coli by transposon mutagenesis and molecular cloning. J Bacteriol 153:408–415

    Google Scholar 

  • Hazelbauer GL, Adler J (1971) Role of the galactose-binding protein in chemotaxis of Escherichia coli toward galactose. Nature New Biol 230:101–104

    Google Scholar 

  • Hazelbauer GL, Harayama S (1983) Sensory transduction in bacterial chemotaxis. Int Rev Cytol 81:33–70

    Google Scholar 

  • Hazelbauer GL, Engstrom P, Harayama S (1981) Methyl-accepting chemotaxis protein III and trasdducer gene trg. J Bacteriol 145:43–49

    Google Scholar 

  • Higgins CF, Hiles ID, Salmond GPC, Gill DR, Downie JA, Evans IJ, Holland IB, Gray L, Buckel SD, Bell AW, Hermondson MA (1986) A family of related ATP-binding subunits coupled to many distinct biological processes in bacteria. Nature 323:448–450

    Google Scholar 

  • Koman A, Harayama S, Hazelbauer GL (1979) Relation of chemotactic response to the amount of receptor: Evidence for different efficiencies of signal transduction. J Bacteriol 138:739–747

    Google Scholar 

  • Kondoh H, Ball CB, Adler J (1979) Identification of a methyl accepting chemotaxis protein for the ribose and galactose chemoreceptors of Escherichia coli. Proc Natl Acad Sci USA 76:260–264

    Google Scholar 

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

    Google Scholar 

  • Lengeler J, Hermann KO, Unsöld HJ, Boos W (1971) The regulation of the β-methylgalactoside transport system of the galactose binding protein of Escherichia coli K12. Eur J Biochem 19:457–470

    Google Scholar 

  • Mahoney WC, Hogg RW, Hermodson MA (1981) The amino acid sequence of the D-galactose-binding protein from Escherichia coli K12. J Biol Chem 256:4350–4356

    Google Scholar 

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

    Google Scholar 

  • Manson MD, Kossmann M (1986) Mutations in tar supress defects in maltose chemotaxis caused by specific malE mutations. J Bacteriol 165:34–40

    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 

  • Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321

    Google Scholar 

  • Middendorf A, Schweizer H, Vreemann J, Boos W (1984) Mapping of markers in the gyrA-his region of Escherichia coli. Mol Gen Genet 197:175–181

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Mowbray SL, Petsko GA (1983) The X-ray structure of the periplasmic galactose-binding protein from Salmonella typhimurium at 3.0 Å resolution. J Biol Chem 258:7991–7997

    Google Scholar 

  • Müller N, Heine HN, Boos W (1985) Characterisation of the Salmonella typhimurium mgl operon and its gene products. J Bacteriol 163:37–45

    Google Scholar 

  • Neu HC, Heppel LA (1965) The release of enzymes of Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 254:7529–7533

    Google Scholar 

  • Ordal GW, Adler J (1974a) Isolation and complementation of mutants in galactose taxis and transport. J Bacteriol 117:509–516

    Google Scholar 

  • Ordal GW, Adler J (1974b) Properties of mutants in galactose taxis and transport. J Bacteriol 117:517–526

    Google Scholar 

  • Randall LL, Josefsson L-G, Hardy SJS (1980) Novel intermediates in the synthesis of maltose-binding protein in Escherichia coli. Eur J Biochem 107:375–379

    Google Scholar 

  • Rotman B, Guzman RJ (1982) Identification of the mglA gene product in the β-methylgalactoside transport system of Escherichia coli. J Biol Chem 257:9030–9034

    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 

  • Scripture JB, Hogg RW (1983) The nucleotide sequences defining the signal peptides of the galactose-binding protein and the arabinose-binding protein. J Biol Chem 258:10853–10855

    Google Scholar 

  • Shuman HA (1981) The use of gene fusions to study bacterial transport proteins. J Membr Biol 61:1–11

    Google Scholar 

  • Tinoco I Jr, Borer PN, Dengler B, Levine M, Uhlenbeck OC, Crothers DM, Gralla J (1973) Improved estimation of secondary structure in ribonucleic acids. Nature New Biol 246:40–41

    Google Scholar 

  • Vyas NK, Vyas MN, Quiocho FA (1983) The 3Å resolution structure of a D-galactose-binding protein for transport and chemotaxis in Escherichia coli. Proc Natl Acad Sci USA 80:1792–1796

    Google Scholar 

  • Zukin RS, Hartig PR, Koshland DE Jr (1977) Use of a distant reporter group as evidence for a conformational change in a sensory receptor. Proc Natl Acad Sci USA 74:1932–1936

    Google Scholar 

Download references

Author information

Author notes

  1. Michael D. Manson

    Present address: Department of Biology, Texas A & M University, 77843, College Station, TX, USA

Authors and Affiliations

  1. Department of Biology, University of Konstanz, D-7750, Konstanz, Federal Republic of Germany

    Annette Scholle, Jörg Vreemann, Volker Blank, Annette Nold, Winfried Boos & Michael D. Manson

Authors
  1. Annette Scholle
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jörg Vreemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Volker Blank
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Annette Nold
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Winfried Boos
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Michael D. Manson
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by J. Lengeler

Rights and permissions

Reprints and permissions

About this article

Cite this article

Scholle, A., Vreemann, J., Blank, V. et al. Sequence of the mglB gene from Escherichia coli K12: Comparison of wild-type and mutant galactose chemoreceptors. Mole Gen Genet 208, 247–253 (1987). https://doi.org/10.1007/BF00330450

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330450

Key words

Search

Navigation