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Nucleic Acids Research, 2003, Vol. 31, No. 16 4682-4688
© 2003 Oxford University Press

Repairing the Sickle Cell mutation. III. Effect of irradiation wavelength on the specificity and type of photoproduct formed by a 3'-terminal psoralen on a third strand directed to the mutant base pair

Steven L. Broitman, Olga Amosova and Jacques R. Fresco*

Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA

*To whom correspondence should be addressed. Tel: +1 609 258 3927; Fax: +1 609 258 2759; Email: jrfresco{at}princeton.edu
Current address:
Steven L. Broitman, Department of Biology, West Chester University, West Chester, PA, USA

Using a psoralen delivery system mediated by a DNA third strand that binds selectively to linear target duplexes immediately downstream from the Sickle Cell ß-globin gene mutation and the comparable wild-type ß-globin gene sequence, the kinetics of formation and yield of psoralen monoadducts and crosslinks with pyrimidine residues at and near the mutant base pair site and its wild-type counterpart were determined. By exploiting irradiation specificities at 300, 365 and 419 nm, it was possible to evaluate the orientation equilibrium of 3'-linked intercalated psoralen and to develop conditions that lead to preferential formation of each type of photoproduct in both the mutant and wild-type sequences. This makes possible the preparation of each type of photoproduct for use as a substrate for DNA repair. In this way, the base pair change(s) that each generates can be established.


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