Urocanic acid photobiology. Photochemical binding to calf thymus DNA

Abstract— Urocanic acid (UA) has previously been shown to photochemically react with N, N-dimethylthymine and with phage G4 single-stranded DNA. In this study, E-ring-2-¹⁴C]-U A and calf thymus DNA have been irradiated with UV light (Λ > 270 nm) in buffered, aqueous solutions. Rc-isolation of the DNA indicates covalent binding of UA at levels of up to 80 nmol UA/mg DNA. Binding is observed for both native and heat denatured DNA. Equilibrium dialysis studies give no evidence for complexation of UA to either form of DNA in the dark. Enzymatic cleavage of the UA bound DNA and analysis by HPLC shows peaks for ^l4C-labelled products with retention volumes identical to those of a marker mixture prepared by irradiating UA with ³H-methyl]thymidine. Photolysis of the DNA before irradiation with U A leads to the formation of a second product with a retention volume corresponding to that for a deoxyadenosine/UA marker.