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Arrest of DNA Elongation by DNA Polymerases at Guanine Adducts on 4-Hydroxyaminoquinoline 1-Oxide-modified DNA Template¹

Department of Biochemistry, Institute for Developmental Research, Aichi Prefecture Colony, Kasugai, Aichi 480-03 [S. Y., O. K., R. S.], and Laboratory of Biochemistry, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya 464 [M. T.], Japan

In vitro modification of M13 phage single-stranded DNA with 4-hydroxyaminoquinoline 1-oxide (4HAQO) resulted in four kinds of adducts: three guanine adducts, QG_I, QG_II, and QG_III; and one adenine adduct, QA, at ratios of 16.4 47.3, 13.7, and 22.6, respectively. The carcinogen-modified DNA, initiated with a sequence-defined oligodeoxynucleotide primer, was replicated in vitro with Escherichia coli DNA polymerase I (Klenow fragment) and calf thymus DNA polymerases and ß. The reaction products were analyzed on a DNA-sequencing gel. DNA elongation by DNA polymerase I was arrested at putative guanine adducts on the template in three ways: at one base prior to guanine; at positions opposite to guanine; and at one base beyond guanine. Similar patterns of elongation arrest were also obtained with the mammalian DNA polymerases and ß. In contrast to guanine adducts, the adenine adduct, QA, might lack the capacity to arrest DNA chain elongation by DNA polymerases.

¹ This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science, and Culture of Japan.

² To whom requests for reprints should be addressed.

Received 9/26/83. Accepted 2/ 3/84.