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Nucleic Acids Research, 1985, Vol. 13, No. 2 319-335
© 1985

Articles

Excision of uracil residues in DNA: mechanism of action of Escherichia coli and Micrococcus luteus uracil-DNA glycosylases

Anne-Marie Delort*, Anne-Marie Duplaa, Didier Molko, Robert Teoule, Jean-Pierre Leblanc and Jacques Laval

Laboratoires de Chimie, Département de Recherche Fondamentale, Centre d'Etudes Nucléaires de Grenoble 85X, F.38041 Grenoble Cedex, France Unité de Biochimie et Enzymologie, Institut Gustave Roussy F.92805 Villejuif Cedex, France

*To whom correspondence should be addressed

Received December 6, 1984. Accepted January 3, 1985.

Various octadeoxynucleotides containing uracil at different positions were synthesized and submitted to the action of Escherichia coli and Micrococcus luteus uracil-DNA glycosylases. A uracil residue situated at the 5'-end was excised by the M.luteus enzyme but not by the E.coli one. Uracil residues located at the ultimate and penultimate positions at the 3'-end were not cleaved by either enzymes. At the other central positions, uracil was eliminated with different initial velocities. Single stranded {varphi}X 174 DNA fragments were used to study the influence of the sequence. Cytosine bases were deaminated to give uracil by bisulfite treatment. It was shown that the initial excision velocity of two vicinal uracil residues was decreased. The same observation was made for two uracils separated by one base. A hypothetical scheme is suggested to explain the mechanism of action of uracil-DNA glycosylases.


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