Bifunctional Photobinding of Psoralen to Single Stranded Nucleic Acids

As psoralen and other furocoumarin derivatives, intercalated between two base pairs of native DNA, under irradiation at 365 nm form inter-strand cross-linkings as a consequence of bifunctional addition, the writers have investigated the ability of psoralen to give such bifunctional photo­ additions, too, with nucleic acids with disordered or partilly disordered structure (denatured DNA and r-RNA). On the basis of fluorimetric, light-scattering, viscosimetric measurements and of the renaturation ability of denatured bacterial DNA, certain results have been obtained. In addition to monofunctional photoadditions, psoralen can give bifunctional binding by irradiation at 365 nm both with denatured DNA and with r-RNA. However, when irradiation of denatured DNA in the presence of psoralen was performed in a concentrated solution (0.4%), the formation of bifunctional additions between two different strands was demonstrated by the increase (50%) of molecular weight of denatured DNA. However, when irradiation of denatured DNA was performed in more dilute solutions (0.1%), the bifunctional photoaddition of psoralen took place producing only bi­ functional additions in the same strand, very probably with the formation of loops, as has been shown by the absence of increase of molecular weight of DNA and by the more restricted structure assumed by the macromolecule, revealed by the light-scattering and viscosimetric measurements. The formation of these bifunctional additions was confirmed by the reduced rate of renaturation shown by denatured bacterial DNA after irradiation in the presence of psoralen. In the case of r-RNA, psoralen, when irradiated can form bifunctional additions only in the same strand.