Effects of base damages on DNA replication. |
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| Authors: | H Ide Y Kimura A Murakami S S Wallace K Makino |
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| Affiliation: | Department of Polymer Science and Engineering, Faculty of Textile Science, Kyoto Institute of Technology, Japan. |
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| Abstract: | Understanding the response of DNA polymerase to the encountered damage in a template is a key to assessing lethal and mutagenic events of cells exposed to genotoxic agents. In the present study M13 (or f1) DNA templates containing 4 types of thymine damages were prepared, and DNA synthesis was carried out in vitro with the templates. The extent of inhibition of DNA synthesis by the damages was evaluated by measuring [3H]dTMP incorporation. Furthermore, newly synthesized DNA was analyzed on a sequencing gel to determine termination sites of DNA synthesis. The results showed that DNA synthesis was differentially inhibited by the damages, and the termination sites of DNA synthesis were dependent on the structures of the damages and the 3'-5' exonuclease activity of DNA polymerase used. |
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