Distamycin analogues without leading amide at their N-termini - comparative binding properties to AT- and GC-rich DNA sequences

Abstract

An efficient, simple and general route towards the solution-phase synthesis of four distamycin analogues containing 2-5 N-methylcarboxamide units without the leading amide unit at the N-terminus is described. The binding abilities of these molecules to calf thymus DNA, poly d(AT), poly dA.poly dT and poly d(GC) were evaluated by duplex DNA melting temperature (T_m) analysis, fluorescence probe displacement assay, footprinting studies and induced circular dichroism (ICD) measurements. A minimum of three N-methylpyrrolecarboxamide units was found to be necessary for the onset of DNA binding. The other three analogues exhibited AT-specific footprints on DNA at a salt concentration of 40 mM NaCl. Interestingly, intense ICD spectra were obtained not only with AT-rich DNA tracts, but also with poly d(GC). Though these ICD signals were sensitive to changes in salt concentration of the solution, residual ICD was present even at [NaCl] values as high as 4.8 M, at which poly d(GC) is likely to exist in the Z conformation. This implies that nonelectrostatic interactions are involved in the binding process involving poly d(GC) and also that binding is preserved even with the Z form of DNA. These results have significance on account of the growing interest in polyamide-based minor groove binders as artificial gene regulators and also in view of the increasing evidence for the biological significance of the Z morph of DNA.