title: Nuclear magnetic resonance study of complexes between CRP and its 22- and 28-base-pair operators.
abstract: The binding of the cAMP receptor protein (CRP) to the portion of the lac promoter comprising the core of the CRP recognition sequence has been investigated. The effect of the binding of CRP to the symmetrical 22- and 28-base-pair operators was investigated by 1H NMR. The binding of cAMP*CRP to the 22mer DNA did not bring about any changes in the chemical shift values of the imino proton resonances of the DNA, but did cause selective line broadening of the imino proton resonances of specific base pairs (TA 4, GC 5). These base pairs are contained in the motif 5'(TGTGA)3', which is thought to be the region crucial to interaction with the CRP. The binding of cAMP*CRP to the 28mer DNA brought about large changes in the imino proton resonances that seem to be induced by DNA bending. Therefore it seems likely that CRP requires DNA longer than a 22mer to bend it. We also used a C-terminal protease-digested CRP (CRPcy) in a DNA-binding experiment. The binding of cAMP*CRPcy to the 28mer DNA did not induce bending. These results indicate that the C-terminal region of CRP participates in DNA binding and is important for DNA bending.
authors.affiliation: Department of Pharmacy, Seoul National University, Korea.
authors.forename: B J
authors.initials: BJ
authors.lastname: Lee
authors.forename: T W
authors.initials: TW
authors.lastname: Lee
authors.forename: S H
authors.initials: SH
authors.lastname: Park
authors.forename: H
authors.initials: H
authors.lastname: Aiba
authors.forename: C
authors.initials: C
authors.lastname: Kojima
authors.forename: Y
authors.initials: Y
authors.lastname: Kyogoku
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D002942:Circular Dichroism; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D002373:Cyclic AMP Receptor Protein; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins