title: DNA bending and twisting properties of integration host factor determined by DNA cyclization.
abstract: The binding of many proteins to DNA is profoundly affected by DNA bending, twisting, and supercoiling. When protein binding alters DNA conformation, interaction between inherent and induced DNA conformation can affect protein binding affinity and specificity. Integration host factor (IHF), a sequence-specific, DNA-binding protein of Escherichia coli, strongly bends the DNA upon binding. To assess the influence of inherent DNA bending on IHF binding, we took advantage of the high degree of natural static curvature associated with an IHF site on a 163-bp minicircle and measured the binding affinity of IHF for its recognition site contained on this DNA in both circular and linear form. IHF showed a higher affinity for the circular form of the DNA when compared to the linear form. In addition, the presence of IHF during DNA cyclization changed the topology of cyclization products and their ability to bind IHF, consistent with IHF untwisting DNA. These results show that inherent DNA conformation anisotropy is an important determinant of IHF binding affinity and suggests a mechanism for modulation of IHF activity by local DNA conformation.
authors.affiliation: University of Southern California, 925 West 34th Street, Los Angeles, California, 90089-0641, USA.
authors.forename: B
authors.initials: B
authors.lastname: Teter
authors.forename: S D
authors.initials: SD
authors.lastname: Goodman
authors.forename: D J
authors.initials: DJ
authors.lastname: Galas
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism*; D004270:DNA, Circular / Q000737:chemistry* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D004996:Ethidium; D034982:Integration Host Factors; D009690:Nucleic Acid Conformation; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D004268:DNA-Binding Proteins; D034982:Integration Host Factors; D004996:Ethidium