© 1995 Oxford University Press
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Induction of mutations by replication of malondialdehyde modified M13 DNA in Escherichia coil: determination of the extent of DNA modification, genetic requirements for mutagenesis, and types of mutations induced
1A.B. Hancock Jr Memorial Laboratory for Cancer Research, Departments of Biochemistry, Chemistry Nashville, TN 37232-0146, USA
2Immunology/Microbiology, Vanderbili University School of Medicine Nashville, TN 37232-0146, USA
3To whom correspondence should be addressed
The mutagenicity of the lipid peroxidatlon product, malondialdehyde (MDA), was measured in the lacZ
forward mutation assay using a recombinant M13 phage, M13MB102. Single-stranded M13MB102 DNA was reacted with MDA at neutral pH and the modified DNA was transformed Into strains of Escherichia coil Induced for the SOS response. Increasing concentrations of MDA led to an increase In lacZ-mutations coincident with an Increase In the level of the major MDA-deoxyguanoslne adduct. Spontaneous and MDA-lnduced M13MB102 mutants were collected and the lacZ target region was subjected to automated DNA sequence analysis. The most common sequence changes induced by MDA were base-pair substitutions (76%). Of these, 43% (29/68) were transverslons, most of which were G
(24/29). Transitions account for 57% of the base-pair substitutions (39/68) and were comprised exclusively of CT (22/39) and A (17/39). Frameshlft mutations were identified In 16% of the induced mutants and were comprised of mainly single base additions occurring In runs of reiterated bases (11/14). The diversity of base-pair substitution and franieshift mutations induced by MDA at low levels of adduction suggests It may be an Important contributor to endogenous mutagenesls and carcinogenesis in aerobic organisms.
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