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DETECTION OF PYRIMIDINE DIMERS AND MONOADDUCTSINDUCED BY 7‐METHYLPYRIDO(3,4‐c) PSORALEN AND UVA IN CHINESE HAMSTER V79 CELLS BY ENZYMATIC CLEAVAGE AND HIGH‐PRESSURE LIQUID CHROMATOGRAPHY
Author(s) -
Dardalhon M.,
Guillo L. A.,
Moysan A.,
Vigny P.,
Averbeck D.
Publication year - 1994
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1994.tb05059.x
Subject(s) - psoralen , chinese hamster , chemistry , cleavage (geology) , enzyme , pyrimidine , high performance liquid chromatography , bond cleavage , chromatography , stereochemistry , biochemistry , dna , biology , catalysis , paleontology , fracture (geology)
The photochemotherapeutically active psoralen derivative 7‐methylpyrido(3,4‐c) psoralen (MePyPs) has been recently shown to be able to photoinduce monoadducts of the C 4 ‐cycloaddition type as well as pyrimidine dimers in DNA in vitro . In the present study, we report on the induction of these two types of photolesions in mammalian cells in culture. The MePyPs photocycloadducts were quantified in V79 Chinese hamster cells after treatment with MePyPs plus UVA following enzymatic hydrolysis of the DNA by DNase I, S1 nuclease and acidic phosphatase treatments. Concomitantly induced pyrimidine dimers were determined by two methods, high‐pressure liquid chromatography and alkaline gel electrophoresis after dimer‐specific endonucleolytic cleavage. The results show that, in Chinese hamster cells treated with MePyPs plus UVA, the yield of pyrimidine dimers is approximately 5‐10% that of MePyPs‐DNA photocycloadducts. Because psoralen monoadditions to DNA alone are generally not considered as being very phototoxic, a synergistic interaction of monoadditions with pyrimidine dimers may be expected to occur in order to explain the high photobiological effectiveness of this psoralen derivative.