Distamycin A affects the stability of NF-κB p50-DNA complexes in a sequence-dependent manner

✍ Scribed by Robert E. Speight; Darren J. Hart; Jonathan M. Blackburn

• Publisher: John Wiley and Sons
• Year: 2002
• Tongue: English
• Weight: 155 KB
• Volume: 15
• Category: Article
• ISSN: 0952-3499
• DOI: 10.1002/jmr.556

✦ Synopsis

Abstract

The effect of two different DNA minor groove binding molecules, Hoechst 33258 and distamycin A, on the binding kinetics of NF‐κB p50 to three different specific DNA sequences was studied at various salt concentrations. Distamycin A was shown to significantly increase the dissociation rate constant of p50 from the sequences PRDII (5′‐GGGAAATTCC‐3′) and Ig‐κ B (5′‐GGGACTTTCC‐3′) but had a negligible effect on the dissociation from the palindromic target‐κ__B__ binding site (5′‐GGGAATTCCC‐3′). By comparison, the effect of Hoechst 33258 on binding of p50 to each sequence was found to be minimal. The dissociation rates for the protein–DNA complexes increased at higher potassium chloride concentrations for the PRDII and Ig‐κ__B__ binding motifs and this effect was magnified by distamycin A. In contrast, p50 bound to the palindromic target‐κ__B__ site with a much higher intrinsic affinity and exhibited a significantly reduced salt dependence of binding over the ionic strength range studied, retaining a K~D~ of less than 10 pM at 150 mM KCl. Our results demonstrate that the DNA binding kinetics of p50 and their salt dependence is strongly sequence‐dependent and, in addition, that the binding of p50 to DNA can be influenced by the addition of minor groove‐binding drugs in a sequence‐dependent manner. Copyright © 2002 John Wiley & Sons, Ltd.