Nucleic Acids Research, Vol 25, Issue 2 304-311, Copyright © 1997 by Oxford University Press
H Kamiya and H Kasai
To evaluate the mutation frequency and the mutation spectrum of 2-
hydroxyadenine (2-OH-Ade), an oxidative DNA lesion, the modified base was
site-specifically incorporated into a unique restriction enzyme site (SalI,
GTCGA*C or AflII, CTTA*AG where A* represents 2-OH-Ade) in single- and
double-stranded vectors. The 2-OH-Ade residues were introduced into (+)-
and (-)-strands of the double-stranded vectors and into the (+)-strand of
single-stranded vectors. When the vectors were transfected intoEscherichia
coli, the modified base showed little to no cytotoxicity. The mutation
frequencies of 2-OH-Ade in the SalI and AflII sites were approximately 0.8
and 0.07%, respectively, with double- stranded (+)-vectors. An increase in
the mutation frequencies was not observed with single-stranded vectors.
When incorporated into the (-)- strand, the mutation frequencies of
2-OH-Ade in the SalI and AflII sites were approximately 0.3 and 0.1%,
respectively. The mutations observed most frequently were -1 deletions at
both positions, in the case of the (+)-strand. On the other hand, we
observed that 2-OH-Ade in the (-)-strand induced A-->G and A-->T
substitutions. These results indicate that 2-OH-Ade residues in DNA induce
substitution and deletion mutations without blocking replication inE.coli.
ARTICLES
Substitution and deletion mutations induced by 2-hydroxyadenine in Escherichia coli: effects of sequence contexts in leading and lagging strands
Department of Environmental Oncology, Institute of Industrial Ecological Sciences, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807, Japan.
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