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Nucleic Acids Research, 1987, Vol. 15, No. 18 7411-7427
© 1987

Articles

Methylene blue photosensitised strand cleavage of DNA: effects of dye binding and oxygen

Colm OhUigin, David J. McConnell, John M. Kelly1 and Wilhelm J. M. van der Putten1,2

1Departments of Genetics and Chemistry, University of Dublin Trinity College, Dublin 2 2Departments of Genetics and School of Physical Sciences, National Institute for Higher Education Glasnevin, Dublin 9, Ireland

Received June 1, 1987. Revised August 14, 1987. Accepted August 21, 1987.

It is shown that methylene blue (MB+) photosensitises DNA in either aerated or deaerated solutions, causing direct cleavage of phosphodiester bonds and rendering additional bonds labile to alkali. Evidence from unwinding and fluorimetric studies indicates that MB+ binds to DNA in at least 2 ways. Intercalation, which optimally induces helical unwinding of 24°±2° per MB+, is markedly reduced upon neutralisation by Mg2+ of the DNA phosphates, while significant non-intercalative binding persists as shown by substantial fluorescence quenching at Mg2+ concentrations where there is little unwinding. MB+ induces photolysis at both low and high Mg2+ concentration - intercalation is apparently not required for photolysis. The quantum yield for strand breakage varies from 1–3 x 10–7 under different conditions and is oxygen enhanced. The DNA cleavage is guanine specific. The 3' termini of the primary MB+-induced DNA photoproducts, unlike those generated by chemical sequencing retain an alkali labile adduct on the terminal phosphate.


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