title: The binding of the cyclic AMP receptor protein to synthetic DNA sites containing permutations in the consensus sequence TGTGA.
abstract: The binding of the cyclic AMP receptor protein (CRP) to symmetrical synthetic DNA-binding sites was investigated with a gel-retardation assay. A set of ten different sequences was employed, comprising all base permutations at positions 2, 4, and 5 of the consensus sequence 5'(TGTGA)3'. We show that: (i) CRP has a higher affinity for the completely symmetrical site than towards the lac wild-type site; (ii) base substitutions at position 2 lead to either a complete loss of specific CRP binding (G----C), a reduction in specific CRP binding (G----A) or only marginal effects on specific CRP binding (G----T); (iii) changes at position 4 abolish (G----C; G----A) or reduce (G----T) specific CRP binding; and (iv) base permutations at position 5 reduce specific CRP binding, but never completely abolish it. Thus position 4, and to a lesser extent position 2, in the DNA consensus sequence are the most crucial ones for specific binding by CRP.
authors: Max-Planck Institut fur Biochemie, Martinsried bei München, Federal Republic of Germany.
authors: C
authors: C
authors: Jansen
authors: A M
authors: AM
authors: Gronenborn
authors: G M
authors: GM
authors: Clore
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein; D004247:DNA / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D009690:Nucleic Acid Conformation; D011953:Receptors, Cyclic AMP / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D046911:Macromolecular Substances; D011953:Receptors, Cyclic AMP; D004247:DNA