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Analysis of the nag regulon from Escberichia coli K12 and Klebsiella pneumoniae and of its regulation

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Summary

Four genes, nagR, A, B and E, clustered in the nag locus of Escherichia coli K12 and Klebsiella pneumoniae, were cloned and physically mapped, and the corresponding gene products involved in amino sugar metabolism identified. Expression of the nag genes was also analysed using a series of lacZ fusions. In both bacteria, the genes are arranged in two divergent operons and controlled by a common NagR repressor. The corresponding gene nagR was found to map in the first operon together with the promoter proximal gene nagB, encoding the enzyme d-glucosamine isomerase (deaminase) (NagB) and the middle gene nagA, coding for N-acetyl-glucosamine deacetylase (NagA). Polar mutations in nagB and nagA prevent the efficient expression of nagR and cause constitutive expression of all nag genes. This includes the gene nagE encoding Enzyme IINag of the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS), encoded in the second divergently transcribed operon. No further gene is found in this operon which in both organisms is directly adjacent to the gene glnS. It is interesting that the NagR repressor also affects the mannose PTS (genes manX, Y, Z), the second transport system involved in amino sugar uptake and phosphorylation.

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References

  • Altenbuchner J, Schmid K, Schmitt R (1983) Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance. J Bacteriol 153:116–123

    Google Scholar 

  • Bachmann BJ (1987) Linkage map of Escherichia coli K12. In: Neidhardt FC, Ingraham JL, Brooks Low K, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium. ASM, Washington DC

    Google Scholar 

  • Bates CJ, Pasternak CA (1965) Further studies on the regulation of amino sugar metabolism. Biochem J 96:147–154

    Google Scholar 

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

    Google Scholar 

  • Bremer E, Silhavy TJ, Weinstock GM (1985) Transposable λplacMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Eschericha coli. J Bacteriol 162:1092–1099

    Google Scholar 

  • Comb DG, Roseman S (1958) Glucosamine-6-phosphate deaminase. Biochim Biophys Acta 21:193–194

    Google Scholar 

  • Ebner R, Lengeler JW (1988) Sucrose-specific Enzyme IISer of the bacterial phosphotransferase system: nucleotide sequence of the gene scrA from pUR400. Mol Microbiol 2:9–17

    Google Scholar 

  • Erni B, Zanolari B (1986) The mannose permease of Escherichia coli consists of three proteins: Amino acid sequence and function in sugar transport, sugar phosphorylation, and penetration of phage DNA. J Biol Chem 262:5238–5247

    Google Scholar 

  • Felton J, Michaelis S, Wright A (1980) Mutations in two unlinked genes are required to produce asparagine auxotrophy in Escherichia coli. J Bacteriol 142:221–228

    Google Scholar 

  • Holmes RP, Russel RRB (1972) Mutations affecting amino sugar metabolism in E. coli K12. J Bacteriol 111:290–291

    Google Scholar 

  • Imada A, Nozaki Y, Kawashima F, Yonida M (1977) Regulation of glucosamine utilization in Staphylococcus aureus and Escherichia coli. J Gen Microbiol 100:329–337

    Google Scholar 

  • Jones-Mortimer MC, Kornberg HL (1980) Amino-sugar transport systems of Escherichia coli K12. J Gen Microbiol 117:369–376

    Google Scholar 

  • Lengeler J (1975) Mutations affecting transport of the hexitols d-mannitol, d-glucitol, and galactitol in Escherichia coli K-12: isolation and mapping. J Bacteriol 124:26–38

    Google Scholar 

  • Lengeler J (1979) Streptozotocin, an antibiotic superior to penicillin in the selection of rare bacterial mutations. FEMS Microbiol Lett 5:417–419

    Google Scholar 

  • Lengeler JW (1980) Characterisation of mutants of Escherichia coli K12, selected by resistance to Streptozotocin. Mol Gen Genet 179:49–54

    Google Scholar 

  • Lengeler J, Lin ECC (1972) Reversal of the mannitol-sorbitol diauxie in Escherichia coli. J Bacteriol 112:840–848

    Google Scholar 

  • Lengeler J, Steinberger H (1978) Analysis of the regulatory mechanisms controlling the synthesis of the hexitol transport systems in Escherichia coli K-12. Mol Gen Genet 164:163–169

    Google Scholar 

  • Levy GA, McAllan A (1959) The N-acetylation and estimation of hexosamines. Biochem J 73:127–132

    Google Scholar 

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

    Google Scholar 

  • Murray NE, Brammar WJ, Murray K (1977) Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet 150:53–61

    Google Scholar 

  • Pardee AB, Prestidge LS (1961) The initial kinetics of enzyme induction. Biochim Biophys Acta 100:591–602

    Google Scholar 

  • Peri KG, Waygood EB (1988) Sequence of cloned Enzyme IIN-acetyl-glucosamne of the phosphoenolpyruvate:N-acetylglucosamine phosphotransferase system of Escherichia coli. Biochemistry 27:6054–6061

    Google Scholar 

  • Plumbridge J (1987) Organisation of the Escherichia coli chromosome between genes glnS and glnU, V. Mol Gen Genet 209:618–620

    Google Scholar 

  • Postma PW, Lengeler JW (1985) Phosphoenolpyruvate: carbohydrate phosphotransferase system of bacteria. Microbiol Rev 49:232–269

    Google Scholar 

  • Rogers JR, Ohgi T, Plumbridge J, Söll D (1988) Nucleotide sequences of the Escherichia coli nagE and nagB genes: the structural genes for the N-acetylglucosamine transport protein of the bacterial phosphoenolpyruvate: sugar phosphotransferase system and for glucosamine-6-phosphate deaminase. Gene 62:197–207

    Google Scholar 

  • Saris PEJ, Palva ET (1987) The ptsL/pell/ptsM (manXYZ) locus consists of three genes involved in mannose uptake in Escherichia coli K-12. FEMS Microbiol Lett 44:371–376

    Google Scholar 

  • Sarvas M (1971) Mutant of Escherichia coli K-12 defective in d-glucosamine biosynthesis. J Bacteriol 105:467–471

    Google Scholar 

  • Shurvinton CE, Lloyd RG, Benson FE, Attfield PV (1984) Genetic analysis and molecular cloning of the Escherichia coli ruv gene. Mol Gen Genet 194:322–329

    Google Scholar 

  • Sprenger G, Vogler A, Lengeler JW (1986) Selection of auxotrophic and carbohydrate-negative mutants in penicillin-resistant Klebsiella pneumoniae by nalidixic acid treatment. FEMS Microbiol Lett 37:299–304

    Google Scholar 

  • Vogler A (1988) Molekulare Untersuchungen zum N-Acetylglucosamin-Stoffwechsel in Enterobakterien. Evolution des Transportsystems und seine Beteiligung an der Chemotaxis. Dissertation Universität Osnabrück

  • Vogler AP, Lengeler JW (1988) Complementation of a truncated membrane-bound Enzyme IINag from Klebsiella pneumoniae with a soluble Enzyme III in Escherichia coli K12. Mol Gen Genet 213:175–178

    Google Scholar 

  • White RJ (1968) Control of amino sugar metabolism in Escherichia coli and isolation of mutants unable to degrade amino sugars. Biochem J 106:847–858

    Google Scholar 

  • White RJ (1970) The role of the phosphoenolpyruvate phosphotransferase system in the transport of N-acetyl-glucosamine by Escherichia coli. Biochem J 118:89–92

    Google Scholar 

  • Wu HC, Wu TC (1971) Isolation and characterization of a glucosamine-requiring mutant of Escherichia coli K-12 defective in glucosamine-6-phosphate synthetase. J Bacteriol 105:455–466

    Google Scholar 

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Communicated by H. Hennecke

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Vogler, A.P., Lengeler, J.W. Analysis of the nag regulon from Escberichia coli K12 and Klebsiella pneumoniae and of its regulation. Molec. Gen. Genet. 219, 97–105 (1989). https://doi.org/10.1007/BF00261163

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  • DOI: https://doi.org/10.1007/BF00261163

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