Wavelength dependence for the photoreversal of a psoralen-DNA cross-link.
We report an action spectrum for the photoreversal of a psoralen cross-link joining two self-complementary DNA oligonucleotides. The cross-link was formed between two thymines (T) on opposite strands of the DNA oligomers and 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT). For comparison, we also present an action spectrum for the photoreversal of the isolated diadduct T-HMT-T. The wavelength dependence for the diadduct photoreversal parallels its absorption spectrum. Both the diadduct and the cross-linked DNA can be photoreversed by exposure to light with wavelengths between 240 and 313 nm. We did not observe photoreversal at 334 nm or above. At least two distinct absorption bands appear to contribute to photoreversal. We measured a quantum yield of 0.16 for photoreversal of the isolated diadduct at wavelengths between 240 and 266 nm. For wavelengths above 280 nm, the quantum yield is 0.30. We also observed a preferential photoreversal at the furan end of the psoralen in the T-HMT-T diadduct. In contrast, the cross-linked DNA oligonucleotides preferentially photoreversed at the pyrone end of the psoralen adduct. The rate constant for photoreversal of the cross-linked DNA is larger than that for the isolated diadduct at wavelengths below 300 nm.
D001483:Base Sequence; D003432:Cross-Linking Reagents; D004247:DNA; D011564:Furocoumarins; D007700:Kinetics; D008433:Mathematics; D009838:Oligodeoxyribonucleotides; D011067:Poly dA-dT; D013056:Spectrophotometry, Ultraviolet; D014307:Trioxsalen / Q000031:analogs & derivatives; D014316:Tritium
D003432:Cross-Linking Reagents; D011564:Furocoumarins; D009838:Oligodeoxyribonucleotides; D014316:Tritium; D011067:Poly dA-dT; C016507:hydroxymethyltrioxsalen; D004247:DNA; D014307:Trioxsalen