Structure of eukaryotic DNA polymerase δ bound to the PCNA clamp while encircling DNA.

Autor:	Zheng F; Structural Biology Program, Van Andel Institute, Grand Rapids, MI 49503., Georgescu RE; DNA Replication Laboratory, The Rockefeller University, New York, NY 10065.; HHMI, The Rockefeller University, New York, NY 10065., Li H; Structural Biology Program, Van Andel Institute, Grand Rapids, MI 49503; Huilin.Li@vai.org odonnel@rockefeller.edu., O'Donnell ME; DNA Replication Laboratory, The Rockefeller University, New York, NY 10065; Huilin.Li@vai.org odonnel@rockefeller.edu.; HHMI, The Rockefeller University, New York, NY 10065.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2020 Dec 01; Vol. 117 (48), pp. 30344-30353. Date of Electronic Publication: 2020 Nov 17.
DOI:	10.1073/pnas.2017637117
Abstrakt:	The DNA polymerase (Pol) δ of Saccharomyces cerevisiae (S.c.) is composed of the catalytic subunit Pol3 along with two regulatory subunits, Pol31 and Pol32. Pol δ binds to proliferating cell nuclear antigen (PCNA) and functions in genome replication, repair, and recombination. Unique among DNA polymerases, the Pol3 catalytic subunit contains a 4Fe-4S cluster that may sense the cellular redox state. Here we report the 3.2-Å cryo-EM structure of S.c. Pol δ in complex with primed DNA, an incoming ddTTP, and the PCNA clamp. Unexpectedly, Pol δ binds only one subunit of the PCNA trimer. This singular yet extensive interaction holds DNA such that the 2-nm-wide DNA threads through the center of the 3-nm interior channel of the clamp without directly contacting the protein. Thus, a water-mediated clamp and DNA interface enables the PCNA clamp to "waterskate" along the duplex with minimum drag. Pol31 and Pol32 are positioned off to the side of the catalytic Pol3-PCNA-DNA axis. We show here that Pol31-Pol32 binds single-stranded DNA that we propose underlies polymerase recycling during lagging strand synthesis, in analogy to Escherichia coli replicase. Interestingly, the 4Fe-4S cluster in the C-terminal CysB domain of Pol3 forms the central interface to Pol31-Pol32, and this strategic location may explain the regulation of the oxidation state on Pol δ activity, possibly useful during cellular oxidative stress. Importantly, human cancer and other disease mutations map to nearly every domain of Pol3, suggesting that all aspects of Pol δ replication are important to human health and disease. Competing Interests: The authors declare no competing interest. (Copyright © 2020 the Author(s). Published by PNAS.)
Databáze:	MEDLINE
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