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Nucleic Acids Research, 1989, Vol. 17, No. 4 1493-1509
© 1989

MOLECULAR BIOLOGY

Action of a mammalian AP-endonuclease on DNAs of defined sequences

Bjørn-Ivar Haukanes*, Dag E. Helland and Kjell Kleppe

Laboratory of Biotechnology and Department of Biochemistry, University of Bergen Bergen, Norway

*To whom correspondence should be addressed at: Laboratory of Biotechnology, PO Box 3152, Arstad, N-5001, Bergen, Norway

Received December 2, 1988. Accepted January 17, 1989.

An apurinic/apyrimidinic (AP) specific endonuclease from mouse plasmacytoma cells (line MPC-11), was observed to cleave apurinic sites in oligonucleotides 9, 11, 12, 39 and 40 nucleotides in length. However, the enzyme failed to cleave AP-sites in two oligonucleotides 7 nucleotides in length. The maximum rates of digestion observed on these short single-stranded DNA ( fragments were approximately 1/30 of the rates observed on double-stranded DNA (dsDNA). In studies using the Maxam-Gilbert DNA sequencing analysis, apurinic sites in purine-rich regions were preferentially cleaved in dsDNA but not in ssDNA indicating that the enzyme has a sequence preference on dsDNA. These results suggest that some sites on DNA might be more efficiently repaired than others.


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