Affiliation:
1. Eppley Institute for Research in Cancer and Allied Diseases, Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198
Abstract
Significance
Despite the important role of human DNA polymerase α (Polα) in genome mutagenesis, there are no structural studies of Polα infidelity. The functional studies are sparse, lack high-resolution approaches, and are performed at a low salt concentration. Here we report the structure of the human Polα catalytic domain in the complex with an incoming deoxycytidine triphosphate (dCTP) and the template:primer containing a T-C mismatch at the growing primer terminus. Pre-steady-state and binding kinetics conducted at a physiological salt concentration revealed that Polα has a remarkably lower affinity to DNA and deoxynucleotide triphosphate (dNTP) than reported previously. Strikingly, we found that the incoming dNTP plays a crucial role in Polα interaction with DNA and in discrimination against a mismatched template:primer. This work is important for understanding the mechanism of Polα infidelity and provides a foundation for future studies.
Funder
HHS | NIH | National Institute of General Medical Sciences
HHS | NIH | National Cancer Institute
Publisher
Proceedings of the National Academy of Sciences
Cited by
9 articles.
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