Summary
Cloning and analysis of the gene nagE encoding Enzyme IINag (EIINag) from Klebsiella pneumoniae revealed strong similarities with the corresponding gene from Escherichia coli K12. Truncated EIINag proteins were generated by inserting a series of Tn1725 transposons into the structural gene; the positions of the insertions were mapped by restriction enzyme analysis, and the activity of the polypeptides determined by in vitro and in vivo tests. Insertions in the region encoding the amino-terminal half of the protein invariably abolished transport and phosphorylation activity, while truncated proteins lacking a C-terminal domain homologous to the soluble Enzyme III (crr gene) could be complemented by this molecule to nearly wild-type activity.
References
Altenbuchner J, Schmid K, Schmitt R (1983) Tn1721-encoded tetracycline resistance: map** of structural and regulatory genes mediating resistance. J Bacteriol 153:116–123
Bramley HF, Kornberg HL (1987) Sequence homologies between proteins of bacterial phosphoenolpyruvate-dependent sugar phosphotransferase system: identification of possible phosphate-carrying histidine residues. Proc Natl Acad Sci USA 84:4770–4780
Ebner R, Lengeler JW (1988) Sucrose-specific Enzyme IIScr of the bacterial phosphotransferase system: nucleotide sequence of the gene scrA from pUR400, Mol Microbiol 2:9–17
Holmes RP, Russel RRB (1972) Mutations affecting amino sugar metabolism in E. coli K12. J Bacteriol 111:290–291
Lengeler JW (1977) Analysis of mutations affecting dissimilation of galactitol (dulcitol) in Escherichia coli K12. Mol Gen Genet 152:83–91
Lengeler JW (1980) Characterisation of mutants of Escherichia coli K12, selected by resistance to streptozotocin. Mol Gen Genet 179:49–54
Lengeler JW, Auburger AM, Pecher A (1981) The phosphoenolpyruvate-dependent carbohydrate: phosphotransferase system enzymes II as chemoreceptors in chemotaxis of Escherichia coli K12. Mol Gen Genet 183:163–170
Lengeler JW, Mayer RJ, Schmid K (1982) Phosphoenolpyruvate-dependent phosphotransferase system Enzyme III and plasmidencoded sucrose transport in Escherichia coli K12. J Bacteriol 151:468–471
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, New York
Plumbridge J (1987) Organisation of the Escherichia coli chromosome between genes glnS and glnU, V. Mol Gen Genet 209:618–620
Postma PW, Lengeler JW (1985) Phosphoenolpyruvate: carbohydrate phosphotransferase system of bacteria. Microbiol Rev 49:856–871
Saier MH, Grenier FC, Lee CA, Waygood EB (1985) Evidence for the evolutionary relatedness of the proteins of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. J Cell Biochem 27:43–56
Sprenger GA, Lengeler JW (1984) L-Sorbose metabolism in Klebsiella pneumoniae and Sor+ derivatives of Escherichia coli K12 and chemotaxis toward sorbose. J Bacteriol 157:39–45
VanGijsegem F, Toussaint A (1982) Chromosome transfer and R-prime formation by an RP4::mini-Mu derivative in Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae and Proteus mirabilis. Plasmid 7:30–44
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Communicated by H. Böhme
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Vogler, A.P., Lengeler, J.W. Complementation of a truncated membrane-bound enzyme IINag from Klebsiella pneumoniae with a soluble enzyme III in Escherichia coli K12. Molec. Gen. Genet. 213, 175–178 (1988). https://doi.org/10.1007/BF00333417
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DOI: https://doi.org/10.1007/BF00333417