Skip to main content
Log in

The mglB sequence of Salmonella typhimurium LT2; promoter analysis by gene fusions and evidence for a divergently oriented gene coding for the mgl repressor

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

Summary

The mglB gene of Salmonella typhimurium LT2 coding for the galactose-binding protein (GBP) was sequenced. We compared the deduced amino acid sequence with the GBP sequence of Escherichia coli K12. The mature proteins differ in only 19 of 309 amino acid residues, corresponding to 94% homology. Analysis of the mglB control region by promoter-probe vectors revealed that two promoters, P1 and P2, constitute the mgl control region (P mgl ). P1 and P2 function in a synergistic way. P1 is the main promoter of the operon; its activity is 20 times the activity of P2. Both promoters are activated by the cyclic adenosine monophosphate catabolite activator protein (cAMP/CAP) complex. While P1 is inactive in the absence of the cAMP/CAP complex, there is residual activity of P2 under these conditions. Studies on the inducibility of the mglBAEC operon using multicopy plasmid promoter-probe vectors were hampered by the titration of the mgl repressor resulting in a partially constitutive expression of the mgl operon. The results indicate that only P1 is responding to induction by D-fucose. A weak promoter, P D , within the P1 region but divergent to it was found. P D is neither stimulated by the cAMP/CAP complex nor by D-fucose. We cloned the gene located downstream to P D and found it to strongly repress the expression of the mgl operon. We termed this gene mglD. The presence of D-fucose abolished the repression caused by the plasmid-encoded mglD gene product.

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.

Similar content being viewed by others

Abbreviations

IPTG:

isopropyl-1-thic-β-D-galatopyranoside

ONPG:

2-nitrophenyl-β-D-galatopyranoside

XG:

5-bromo-4-chloro-3-indolyl-β-D-galatopyranoside

Kanr :

Kanamycin resistance

References

  • Anraku Y (1968) Transport of sugars and amino acids in bacteria. J Biol Chem 243:3116–3128

    Google Scholar 

  • Benner D, Müller N, Boos W (1985) Temperature-sensitive catabolite activator protein in Escherichia coli BUG6. J Bacteriol 161:347–352

    Google Scholar 

  • Bolivar F, Rodriguez RL, Green PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction and characterization 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 E. coli. Eur J Biochem 10:66–73

    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 

  • Bremer E, Silhavy TJ, Weisemann JM, Weinstock GM (1984) placMu: a transposible derivative of bacteriophage lambda 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 promoters in E. coli using bacteriophage lambda and Mu. J Mol Biol 104:541–555

    Google Scholar 

  • Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory, Press, Cold Spring Harbor, New York

    Google Scholar 

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

    Google Scholar 

  • Elledge SJ, Walker GC (1985) Phasmid vectors for identification of genes by complementation of E. coli mutants. J Bacteriol 162:777–783

    Google Scholar 

  • Ganesan AK, Rotman B (1965) Transport systems for galactose and galactosides in E. coli. J Mol Biol 16:42–50

    Google Scholar 

  • Griggs DW, Tharp BB, Konisky J (1987) Cloning and promoter identification of the iron-regulated cir gene of E. coli. J Bacteriol 169:5343–5352

    Google Scholar 

  • Harayama S, Bollinger J, Iino T, Hazelbauer GL (1983) Characterization of the mgl operon of E. 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 E. coli toward galactose. Nature (London) New Biol 230:101–104

    Google Scholar 

  • Hogg RW, Isihara H, Hermodson, MA, Koshland D Jr, Jacobs JW, Bradshaw RA (1977) A comparision of the amino-terminal sequences of several carbohydrate binding proteins from E. coli and S. typhimurium. FEBS Lett 80:377–379

    Google Scholar 

  • Jordan E, Saedler H, Lengeler J, Starlinger P (1967) Changes in the specific activities of the galactose enzymes in E. coli under different growth conditions. Mol Gen Genet 100:203–209

    Google Scholar 

  • Lengeler J, Hermann KO, Unsöld HJ, Boos W (1971) The regulation of the β-methylgalactoside transport system and of the galactose binding protein of E. coli K 12. Eur J Biochem 19:457–470

    Google Scholar 

  • Lopilato J, Bortner S, Beckwith, J (1986) Mutations in a new chromosomal gene of E. coli K 12, pcn reduce plasmid copy number of pBR322 and its derivatives. Mol Gen Genet 205:285–290

    Google Scholar 

  • Mahoney WC, Hoog RW, Hermodson MA (1981) The amino acid sequences of the D-galactose-binding protein from E. coli B/r. J Biol Chem 256:4350–4356

    Google Scholar 

  • Majumdar A, Adhya S (1987) Probing the structure of gal operator-repressor complexes. J Biol Chem 262:13258–13262

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Müller N, Heine H-G, Boos W (1982) Cloning of mglB, the structural gene for the galactose-binding protein, of S. typhimurium and E. coli. Mol Gen Genet 185:473–480

    Google Scholar 

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

    Google Scholar 

  • Norrander J, Kempe T, Messing J (1983) Construction of improved M 13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26:101–106

    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 

  • Postma PW (1977) Galactose transport in Salmonella typhimurium. J Bacteriol 129:630–639

    Google Scholar 

  • Ptashne M (1986) Gene regulation by proteins acting nearby and at a distance. Nature 322:697–701

    Google Scholar 

  • Quiocho FA, Pflugrath JW (1980) The structure of D-galactose binding protein at 4.1 Å resolution looks like L-arabinose-binding protein. J Biol Chem 255:6559–6561

    Google Scholar 

  • Raibaud O, Schwartz M (1980) Map of the E. coli malA region and identification of the malT product. J Bacteriol 143:761–771

    Google Scholar 

  • Robbins AR (1975) Regulation of the E. coli β-methylgalactoside transport system by gene mglD. J Bacteriol 123:69–74

    Google Scholar 

  • Rotman B, Guzman RJ (1982) Identification of the mglA gene product in the β-methylgalactoside transport system of E. 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 

  • Schneider K, Beck CF (1986) Promoter-probe vectors for the analysis of divergently arranged promoters. Gene 42:37–48

    Google Scholar 

  • Scholle A, Vreemann J, Blank V, Nold A, Boos W, Manson, MD (1987) Sequence of the mglB gene from E. coli K 12: comparision of the wild-type and mutant galactose chemoreceptors. Mol Gen Genet 208:247–253

    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 

  • Silhavy TJ, Berman ML, Enquist W (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Valentin-Hansen P, Albrechtsen B, Løve Larsen JE (1986) DNA-protein recognition: demonstration of three genetically separated operator elements that are required for repression of the E. coli deoCABD promoters by the DeoR repressor. EMBO J 5:2015–2021

    Google Scholar 

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

    Google Scholar 

  • Vyas NK, Vyas MN, Quiocho FA (1987) A novel calcium binding site in the galactose-binding protein of bacterial transport and chemotaxis. Nature 327:635–638

    Google Scholar 

  • Wilcox G, Boulter J, Lee N (1974) Direction of transcription of the regulatory gene araC in E. coli B/r. Proc Natl Acad Sci USA 71:3635–3639

    Google Scholar 

  • Wu HCP, Kalckar HM (1966) Endogenous induction of the galactose operon in Escherichia coli K 12. Proc Natl Acad Sci USA 55:622–629

    Google Scholar 

  • Zukin RS, Strange PG, Heavey LR, Koshland DE Jr (1977) Properties of the galactose binding protein of S. typhimurium and E. coli. Biochem 16:381–386

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Lengeler

Rights and permissions

Reprints and permissions

About this article

Cite this article

Benner-Luger, D., Boos, W. The mglB sequence of Salmonella typhimurium LT2; promoter analysis by gene fusions and evidence for a divergently oriented gene coding for the mgl repressor. Mol Gen Genet 214, 579–587 (1988). https://doi.org/10.1007/BF00330498

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330498

Key words

Navigation