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Nucleic Acids Research, 1981, Vol. 9, No. 9 2207-2222
© 1981

CHEMISTRY

Identification of the modified nucleotides produced by covalent photoaddition of hydroxy-methyltrimethylpsoralen to RNA

Jean-Pierre Bachellerie1, John F. Thompson, Maurice R. Wegnez2 and John E. Hearst

Department of Chemistry and Laboratory of Chemical Biodynamics, University of California at Berkeley Berkeley, CA 94720, USA

Received January 30, 1981.

The reaction between RNA and 4'hydroxymethyl-4,5', been 8-trimethylpsoralen has been studied. Both natural RNA and synthetic RNAs were used. The base specificity of the reaction was found to be the same in natural RNA, homopolymers, and mononucleotides. Uridine was found to be the most reactive base in all cases. The kinetics of formation and reversal of monoadducts and crosslinks has been examined. Paper electrophoretic conditions are described which provide a separation of the monoaddition and crosslinked photoproducts. The relative and absolute amounts of monoadducts and crosslinks can be determined very accurately with this system. Paper electrophoresis provides good separations of the different photoproducts. The mobilities of the products are a simple function of their molecular weights and charges.

1Present address: Centre de Recherches en Biochimie et Genetique Cellulaires du C.N.R.S., 118, Route de Narbonne, 31077 Toulouse, France

2Present address: Centre de Genetique Moleculaire, C.N.R.S. F-91190, Gif-sur-Yvette, France


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