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J Biol Chem, Vol. 273, Issue 50, 33406-33413, December 11, 1998
From the Donner Laboratory, Lawrence Berkeley National Laboratory,
University of California, Berkeley, California 94720
Previous experiments on DNA sequence context
reported that base modification, replication, and repair are affected
by the nature of neighbor bases. We now report that repair by mammalian alkylpurine-DNA-N-glycosylases (APNG) of 15-mer
oligonucleotides with a central
1,N6-ethenoadenine (
Correlation between Sequence-dependent Glycosylase
Repair and the Thermal Stability of Oligonucleotide Duplexes
Containing 1,N6-Ethenoadenine
A), flanked by 5'
and 3' tandem bases, is also highly sequence dependent.
Oligonucleotides with the central sequences -GGAGG- or -CCACC-
are repaired 3-5-fold more efficiently than those containing
-AAAAA- or -TTATT- when using human or mouse APNG. Melting curves
of the same duplexes showed that oligomers with G·C/C·G neighbors
were less denatured than those with A·T/T·A neighbors at 37 °C.
This sequence-dependent difference in denaturation correlates with the relative thermodynamic stability of oligomers with
G·C/C·G or A·T/T·A neighbors. The dependence of repair on thermal stability was confirmed by enzyme reactions performed over
0-45 °C. Under these conditions, repair of A flanked by G·C/C·G was dramatically increased at 37 °C with continuous
increase up to 45 °C, in contrast to that with flanking A·T/T·A
pairs, which was in agreement with the degree of denaturation of these duplexes. These results indicate that the thermodynamic stability conferred by base pairs flanking A plays an essential role in maintaining the integrity of the duplex structure which is necessary for repair.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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