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Nucleic Acids Research, 1986, Vol. 14, No. 3 1417-1426
© 1986

Articles

Base substitution mutations induced in the cI gene of lambda phage by neocarzinostatin chromophore: correlation with depyrimidination hotspots at the sequence AGC

Lawrence F. Povirk1,2 and Irving H. Goldberg2

1Department of Pharmacology and Toxicology, Medical College of Virginia, Virginia Commonwealth University Richmond, VA 23298 2Department of Pharmacology, Harvard Medical School Boston, MA 02115, USA

Received November 25, 1985. Revised January 13, 1986. Accepted January 13, 1986.

Treatment of intact lambda phage with the nonprotein chromophore of neocarzinostatin resulted in efficient phage inactivation and generation of clear-plaque mutants. Both effects required a preincubation at low pH to allow diffusion of chromophore into the phage head. Chromophore activation was then effected by addition of a sulfhydryl cofactor, followed by a shift to neutral pH. Sequence analysis of mutations mapped to the DNA-binding region of the cl gene revealed that nearly all were single base substitutions. Significant numbers of all possible base changes were found, with A:T to C:C transitions being the most frequent events. Of 11 C:C to A:T transitions, 7 were found at C residues in the trinucleotide sequence AGC, which has previously been shown to be a hotspot for chromophore-induced depyrimidination. This result, as well as the SOS dependence of mutagenesis and the overall distribution of various types of base substitutions, is consistent with the hypothesis that apurinic/apyrimidinic sites are important mutagenic lesions.


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