The structure of the nuclear factor-kappaB protein-DNA complex varies with DNA-binding site sequence.
Transcriptional regulation of many immune responsive genes is under the control of the transcription factor NF-kappaB. This factor is found in cells as a dimer which can contain any two members of the Rel family of proteins (p50, p65, p52, c-Rel, and RelB). The different dimers show distinct preferences for DNA-binding site sequences. To understand the relationship between the DNA binding properties of the dimer forms and transcriptional activation, the physical properties of the complexes of p50 and p65 with DNA have been analyzed. Comparison of apparent DNA binding affinity showed differences in selectivity of DNA-binding site sequence. The ionic strength dependence of apparent binding affinity has shown that the number of ionic interactions in the protein-DNA complex depends on the DNA-binding site sequence and the dimer form, which are consistent with changes in the structure of the protein-DNA complex. Using a fluorescent technique to measure DNA structure changes, protein binding does not appear to alter the structure of the DNA-binding site within the limits of detection. These results are consistent with a change in protein structure that may result in activation differences due to alternative interactions with other transcription proteins.
Menetski
J P
JP
Department of Molecular Biology, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert, Ann Arbor, Michigan 48105, USA. joseph.menetski@wl.com
D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D019281:Dimerization; D004735:Energy Transfer; D005454:Fluorescence Polarization; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D016899:Interferon-beta / Q000235:genetics; D008956:Models, Chemical; D016328:NF-kappa B / Q000737:chemistry* / Q000378:metabolism; D052002:NF-kappa B p50 Subunit; D009690:Nucleic Acid Conformation; D009698:Nucleoproteins / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D051996:Transcription Factor RelA
D007145:Immunoglobulin kappa-Chains; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D009698:Nucleoproteins; D009838:Oligodeoxyribonucleotides; D051996:Transcription Factor RelA; D016899:Interferon-beta; D004247:DNA