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A Redox Role for the [4Fe4S] Cluster of Yeast DNA Polymerase δ

Bartels, Phillip L. and Stodola, Joseph L. and Burgers, Peter M. J. and Barton, Jacqueline K. (2017) A Redox Role for the [4Fe4S] Cluster of Yeast DNA Polymerase δ. Journal of the American Chemical Society, 139 (50). pp. 18339-18348. ISSN 0002-7863. PMCID PMC5881389. doi:10.1021/jacs.7b10284. https://resolver.caltech.edu/CaltechAUTHORS:20171211-090046935

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Abstract

A [4Fe4S]^(2+) cluster in the C-terminal domain of the catalytic subunit of the eukaryotic B-family DNA polymerases is essential for the formation of active multi-subunit complexes. Here we use a combination of electrochemical and biochemical methods to assess the redox activity of the [4Fe4S]^(2+) cluster in Saccharomyces cerevisiae polymerase (Pol) δ, the lagging strand DNA polymerase. We find that Pol δ bound to DNA is indeed redox-active at physiological potentials, generating a DNA-mediated signal electrochemically with a midpoint potential of 113 ± 5 mV versus NHE. Moreover, biochemical assays following electrochemical oxidation of Pol δ reveal a significant slowing of DNA synthesis that can be fully reversed by reduction of the oxidized form. A similar result is apparent with photooxidation using a DNA-tethered anthraquinone. These results demonstrate that the [4Fe4S] cluster in Pol δ can act as a redox switch for activity, and we propose that this switch can provide a rapid and reversible way to respond to replication stress.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/jacs.7b10284DOIArticle
https://pubs.acs.org/doi/suppl/10.1021/jacs.7b10284PublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881389PubMed CentralArticle
ORCID:
AuthorORCID
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2017 American Chemical Society. Received: September 26, 2017; Published: November 22, 2017. We are grateful to the NIH (GM120087 to J.K.B., GM118129 to P.M.B.) and US-Israel Binational Science Foundation (2013358 to P.M.B.) for their financial support of this work. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
NIHGM120087
NIHGM118129
Binational Science Foundation (USA-Israel)2013358
Issue or Number:50
PubMed Central ID:PMC5881389
DOI:10.1021/jacs.7b10284
Record Number:CaltechAUTHORS:20171211-090046935
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171211-090046935
Official Citation:A Redox Role for the [4Fe4S] Cluster of Yeast DNA Polymerase δ Phillip L. Bartels, Joseph L. Stodola, Peter M. J. Burgers, and Jacqueline K. Barton. Journal of the American Chemical Society 2017 139 (50), 18339-18348. DOI: 10.1021/jacs.7b10284
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83788
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:12 Dec 2017 22:30
Last Modified:15 Nov 2021 20:14

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