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J Am Chem Soc. 2013 May 1;135(17):6384-7. doi: 10.1021/ja311434s. Epub 2013 Apr 17.

Tolerance of base pair size and shape in postlesion DNA synthesis.

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Department of Health Sciences and Technology, Institute of Food Nutrition and Health, ETH Zürich, Schmelzbergstrasse 9, 8092 Zürich, Switzerland.


The influence of base pair size and shape on the fidelity of DNA polymerase-mediated extension past lesion-containing mispairs was examined. Primer extension analysis was performed with synthetic nucleosides paired opposite the pro-mutagenic DNA lesion O(6)-benzylguanine (O(6)-BnG). These data indicate that the error-prone DNA polymerase IV (Dpo4) inefficiently extended past the larger Peri:O(6)-BnG base pair, and in contrast, error-free extension was observed for the smaller BIM:O(6)-BnG base pair. Steady-state kinetic analysis revealed that Dpo4 catalytic efficiency was strongly influenced by the primer:template base pair. Compared to the C:G pair, a 1.9- and 79,000-fold reduction in Dpo4 efficiency was observed for terminal C:O(6)-BnG and BIM:G base pairs respectively. These results demonstrate the impact of geometrical size and shape on polymerase-mediated mispair extension.

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