cAMP-cAMP receptor protein complex: five binding sites in the control region of the Escherichia coli mannitol operon.
The control region of the mannitol (mtl) operon of Escherichia coli has been shown to contain five cAMP receptor protein (CRP) binding sequences, the most yet reported for any operon. A DNA fragment encompassing the entire mtl operon regulatory region was generated by PCR, and the binding of the cAMP-CRP complex was studied. Using restrictional analysis to separate, delineate and destroy the various putative CRP binding sites, all five sites were shown to be functional for CRP binding in vitro. Four of these sites bound the cAMP-CRP complex with high affinity while the fifth site (the most distal relative to the transcriptional start site) bound the complex with lower affinity. Simultaneous binding of cAMP-CRP complexes to several of these sites was demonstrated. The results serve to identify and define five dissimilar CRP binding sites in a single operon of E. coli. A model for mtl operon transcriptional initiation and repression complexes is presented.
Ramseier
Tom M
TM
Saier
Milton H
MH
D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D000242:Cyclic AMP / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D008353:Mannitol / Q000378:metabolism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D009876:Operon / Q000235:genetics*; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D002373:Cyclic AMP Receptor Protein; D029968:Escherichia coli Proteins; D009004:Monosaccharide Transport Proteins; D008353:Mannitol; D000242:Cyclic AMP; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C022358:mannitol PTS permease, E coli