Search

The Authors have studied the capacity to photoreact with native DNA and r-RNA using six coumarins and four 4′,5′-dihydro-furo-coumarins. Six of these substances have shown a definite photoreactivity both with DNA and r-RNA, even if in a very much lower extent in respect to that of skin-photosensitizing furocoumarins.

The Authors have studied the biosynthesis of some furocoumarins contained in Ruta graveolens. Labeled 5,7- and 7,8-dihydroxycoumarins have been administered to the herb, however practically no incorporation of radioactivity was observed in the isolated furocoumarins. Moreover, with the aim to study in a more detailed way the role of rutaretin and marmesin, trapping experiments have been performed administering to the herb labeled umbelliferone together with unlabeled rutaretin and marmesin respectively. A very effective incorporation was observed into marmesin, in lesser extent into rutaretin.

A furocoumarin intercalated between two base pairs of native DNA can photoreact with two pyrimidine bases engaging both its 4',5'- and its 3,4-double bond. This fact was evidenced studying the behaviour of the fluorescence acquired by DNA during irradiation at 365 nm. When this double reaction occurs, a cross-linking is formed between the two strands of DNA, as denaturation and renaturation experiments have demonstrated. The various furocoumarin derivatives have a very diverse ability to form cross-linkages, which is independent of their photobinding capacity. This different ability is due mainly to the structure of furocoumarins and to the steric relationships that the intercalated furocoumarin molecules have with the stacked pyrimidine bases.

The photoreaction at 3655 A between skin-photosensitizing furocoumarins and DNA leads to a photo-C4-cyclo-addition reaction in which the 5 - 6 double bond of the pyrimidine bases and the 3 -4 or the 4' -5′ double bond of the furocoumarins are involved. In continuing the research in this field, the authors have investigated the photoreactions (3655 A) of psoralen, xanthotoxin (8-methoxy-psoralen), bergapten (5-methoxy-psoralen) and 8-methyl-psoralen with native calf thymus DNA. with heat-denatured DNA and with ribosomal yeast-RNA, using labelled furocoumarins and examining also the influence of the temperature at which the irradiation was made. At room temperature (22° -30°C) psoralen photoreacts in almost equally extent with each of the three nucleic acid samples, while xanthotoxin, bergapten and 8-methyl-psoralen photoreact to a greater extent with native DNA than with denatured DNA and RNA.

Light scattering measurements performed on DNA after irradiation in the presence of psoralen clearly show that inter strand cross linkings are present in the macromolecule. In fact after heat denaturation and successive cooling irradiated macromolecule shows a molecular weight practically unchanged while a DNA sample after the same treatment shows a molecular weight half of the intact native DNA. Also the general conformation of irradiated DNA undergoes practically to no modifications after the same heat treatment while native DNA shows itself to have been strongly modified. Moreover, on the basis of flow dichroism determinations, DNA cross-linked by psoralen after heat denaturation showed to be able to restore its ordered double helix structure, during the successive cooling.

The biosynthesis of some furocoumarins present in Ruta graveolens has been studied administering labeled psoralen, xanthotoxin, rutaretin, marmesin and 7-hydroxycoumarin-derivatives.

With the aim of studying the biosynthesis of psoralen and bergapten, two furocoumarins present in the leaves of “Ficus carica” (Moraceae), the Authors, continuing preceding researchs on this topic, have fed the leaves with the following labelled precursors: 4′,5′-dihydropsoralen, 4′,5′-dihydrobergapten, 7-hydroxycoumarin, 5,7-dihydroxycoumarin and 5-methoxy-7-hydroxycoumarin.