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Nucleic Acids Research, Vol 26, Issue 19 4432-4438, Copyright © 1998 by Oxford University Press

ARTICLES

An apyrimidinic site kinks DNA and triggers incision by endonuclease VII of phage T4

B Greger and B Kemper
Institute for Genetics of the University of Cologne, Zulpicher Strasse 47, 50674 Koln, Germany.

Apurinic/apyrimidinic lesions (AP-sites) occur frequently in DNA, generated by physically and chemically induced or spontaneous loss of bases. Repair mechanisms have evolved in organisms to deal efficiently with AP-sites by first incising the DNA at the lesion, followed by excision and resynthesis of the damaged strand. Here we report that endonuclease VII (endo VII) of phage T4, which was originally classified as a debranching and Holliday structure resolving enzyme, also recognizes AP-sites with high efficiency. The enzyme cleaves both strands of double-stranded DNA in a stepwise fashion a few nucleotides 3' of the lesion. In a search for a recognition signal shared by all known endo VII substrates, kinking of DNA has earlier been suggested as such a signal. In support of this hypothesis, we demonstrate here that AP-sites induce distinct kinks in synthetic oligonucleotides allowing efficient intramolecular ring closure by ligation.
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