DNA analogues are currently being intensely investigated owing to their potential as gene-targeted drugs. Furthermore, their properties and interaction with DNA and RNA could provide a better understanding of the structural features of natural DNA that determine its unique chemical, biological and genetic properties. We recently designed a DNA analogue, PNA, in which the backbone is structurally homomorphous with the deoxyribose backbone and consists of N-(2-aminoethyl)glycine units to which the nucleobases are attached. We showed that PNA oligomers containing solely thymine and cytosine can hybridize to complementary oligonucleotides, presumably by forming Watson-Crick-Hoogsteen (PNA)2-DNA triplexes, which are much more stable than the corresponding DNA-DNA duplexes, and bind to double-stranded DNA by strand displacement. We report here that PNA containing all four natural nucleobases hybridizes to complementary oligonucleotides obeying the Watson-Crick base-pairing rules, and thus is a true DNA mimic in terms of base-pair recognition.

Linear dichroism (LD) spectroscopy and steady-state as well as time-resolved luminescence spectroscopy have been used to investigate the interaction of the and enantiomers of Ru(phen)2DPPZ2+ (phen = 1,10-phenanthroline; DPPZ = dipyrido[3,2-n:2',3'-c]phenazine) with DNA. The pure enantiomers, which were difficult to separate by traditional resolving methods, were synthesized via a chiral precursor. Changes in luminescence, isotropic absorption and excited state lifetimes upon binding, and the LD observed in flow-oriented DNA systems provide detailed information about the DNA binding of the enantiomers. Flow LD shows that both enantiomers bind to DNA in a well-defined manner with an orientation of the dipyridophenazine chromophore consistent with intercalation of this moiety between base-pairs. Both enantiomers are found to show luminescence in the presence of DNA to which they bind very strongly (K = 10s M™1); however, the relative luminescence quantum yield of the bound enantiomer is 6-10 times larger than that of the bound enantiomer. Furthermore, for each enantiomer two distinct excited state lifetimes are found in varying proportions depending on the binding ratio. The large difference in luminescence quantum yield between the enantiomers is interpreted in terms of slightly different intercalation geometries of the dipyridophenazine ligand, resulting in different protections from quenching by solvent water and diastereomeric differences in the interactions between enantiomers bound in contigue on DNA.

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This review will consider solution studies of structure and interactions of DNA and DNA complexes using linear dichroism spectroscopy, with emphasis on the technique of orientation by flow. The theoretical and experimental background to be given may serve, in addition, as a general introduction into the state of the art of linear dichroism spectroscopy, particularly as it is applied to biophysical problems.

Although the importance of the nucleobases in the DNA double helix is well understood, the evolutionary significance of the deoxyribose phosphate backbone and the contribution of this chemical entity to the overall helical structure and stability of the double helix is not so clear. Peptide nucleic acid (PNA) is a DNA analogue with a backbone consisting of N-(2-aminoethyl)glycine units (Fig. 1) which has been shown to mimic DNA in forming Watson-Crick complementary duplexes with normal DNA. Using circular dichroism spectroscopy we show here that two complementary PNA strands can hybridize to one another to form a helical duplex. There is a seeding of preferred chirality which is induced by the presence of an L- (or D-) lysine residue attached at the carboxy terminus of the PNA strand. These results indicate that a (deoxy)ribose phosphate backbone is not an essential requirement for the formation of double helical DNA-like structures in solution.