Regulation of gluconeogenesis by the glucitol enzyme III of the phosphotransferase system in Escherichia coli.
The gut operon was subcloned into various plasmid vectors (M. Yamada and M. H. Saier, Jr., J. Bacteriol. 169:2990-2994, 1987). Constitutive expression of the plasmid-encoded operon prevented utilization of alanine and Krebs cycle intermediates when they were provided as sole sources of carbon for growth. Expression of the gutB gene alone (encoding the glucitol enzyme III), subcloned downstream from either the lactose promoter or the tetracycline resistance promoter, inhibited utilization of the same compounds. On the other hand, overexpression of the gutA gene (encoding the glucitol enzyme II) inhibited the utilization of a variety of sugars as well as alanine and Krebs cycle intermediates by an apparently distinct mechanism. Phosphoenolpyruvate carboxykinase activity was greatly reduced in cells expressing high levels of the cloned gutB gene but was nearly normal in cells expressing high levels of the gutA gene. A chromosomal mutation in the gutR gene, which gave rise to constitutive expression of the chromosomal gut operon, also gave rise to growth inhibition on gluconeogenic substrates as well as reduced phosphoenolpyruvate carboxykinase activity. Phosphoenolpyruvate synthase activity in general varied in parallel with that of phosphoenolpyruvate carboxykinase. These results suggest that high-level expression of the glucitol enzyme III of the phosphotransferase system can negatively regulate gluconeogenesis by repression or inhibition of the two key gluconeogenic enzymes, phosphoenolpyruvate carboxykinase and phosphoenolpyruvate synthase.
Yamada
M
M
Department of Biology, University of California at San Diego, La Jolla 92093.
Feucht
B U
BU
Saier
M H
MH
D000409:Alanine / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005943:Gluconeogenesis; D026901:Membrane Transport Proteins / Q000096:biosynthesis; D009004:Monosaccharide Transport Proteins; D009154:Mutation; D009876:Operon; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D010957:Plasmids; D011772:Pyruvate, Orthophosphate Dikinase / Q000378:metabolism; D027981:Symporters
D029968:Escherichia coli Proteins; C050221:LacY protein, E coli; D026901:Membrane Transport Proteins; D009004:Monosaccharide Transport Proteins; D027981:Symporters; C021499:lactose permease; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C040707:srlB protein, E coli; D011772:Pyruvate, Orthophosphate Dikinase; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D000409:Alanine