TY - JOUR
T1 - A Redox Role for the [4Fe4S] Cluster of Yeast DNA Polymerase δ
AU - Bartels, Phillip L.
AU - Stodola, Joseph L.
AU - Burgers, Peter M.J.
AU - Barton, Jacqueline K.
N1 - Funding Information:
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.
Publisher Copyright:
© 2017 American Chemical Society.
PY - 2017/12/20
Y1 - 2017/12/20
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85038633721&partnerID=8YFLogxK
U2 - 10.1021/jacs.7b10284
DO - 10.1021/jacs.7b10284
M3 - Article
C2 - 29166001
AN - SCOPUS:85038633721
VL - 139
SP - 18339
EP - 18348
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 50
ER -