Wednesday, 10 November 2004 - 2:40 PM
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This presentation is part of: Nucleic Acids: Structural Motifs and Applications I

How polymerases make mistakes

Gerald Hsu, Sean J. Johnson, Lawrence J. Forsberg, Joshua J. Warren, and Lorena S. Beese. Duke University Medical Center, Durham, NC

Accurate DNA replication is essential for genomic stability. One mechanism by which high-fidelity DNA polymerases maintain replication accuracy involves stalling of the polymerase in response to covalent incorporation of mismatched base pairs, thereby favoring subsequent mismatch excision. Some polymerases retain a "short-term memory" of replication errors, responding to mismatches up to four base pairs in from the primer terminus. We recently presented a structural characterization of all 12 possible mismatches captured at the growing primer terminus in the active site of a polymerase [Johnson & Beese, Cell 116, 803]. Our observations suggest four mechanisms that lead to mismatch-induced stalling of the polymerase. Furthermore, we have observed the effects of extending a mismatch up to six base pairs from the primer terminus and we find that long-range distortions in the DNA transmit the presence of the mismatch back to the enzyme active site, suggesting the structural basis for the short-term memory of replication errors.

By contrast with the 12 naturally occurring base-base mismatches, some DNA damage lesions form DNA mispairs that are efficiently extended. Other recent work from our lab [Hsu et al. & Beese, Nature, 431(7005):217-221] demonstrates how one class of lesions inverts the mechanisms of mismatch recognition, leading to recognition of correctly paired nucleotides as mispairs, and vice versa.


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