Published February 24, 2017 | Version Supplemental Material + Accepted Version
Journal Article Open

The [4Fe4S] cluster of human DNA primase functions as a redox switch using DNA charge transport

Abstract

DNA charge transport chemistry offers a means of long-range, rapid redox signaling. We demonstrate that the [4Fe4S] cluster in human DNA primase can make use of this chemistry to coordinate the first steps of DNA synthesis. Using DNA electrochemistry, we found that a change in oxidation state of the [4Fe4S] cluster acts as a switch for DNA binding. Single-atom mutations that inhibit this charge transfer hinder primase initiation without affecting primase structure or polymerization. Generating a single base mismatch in the growing primer duplex, which attenuates DNA charge transport, inhibits primer truncation. Thus, redox signaling by [4Fe4S] clusters using DNA charge transport regulates primase binding to DNA and illustrates chemistry that may efficiently drive substrate handoff between polymerases during DNA replication.

Additional Information

© 2017 American Association for the Advancement of Science. 19 May 2016; accepted 23 January 2017. Supported by NIH operating grants GM61077 and GM120087 (J.K.B.); NIH operating grants GM65484 and GM118089 (W.J.C.); NIH training grants T32 GM07616 (E.O'B.), T32 GM08230 (M.E.H. and L.E.S.), and T32 CA009582 (A.C.E.); NIH grant S10 RR025677 in support of NMR instrumentation; and the Moore Foundation. Crystallographic data for the p58C Y345F and Y347F mutants have been deposited in the Protein Data Bank under accession numbers PDB 517M and PDB 5DQO, respectively.

Attached Files

Accepted Version - nihms848302.pdf

Supplemental Material - 23/355.6327.eaag1789.DC1/OBrien.SM.pdf

Files

nihms848302.pdf

Files (1.8 MB)

Name Size
md5:350841f2e776677f69e51f9649bb40e2
1.8 MB Preview Download

Additional details

Identifiers

PMCID
PMC5338353
Eprint ID
74510
Resolver ID
CaltechAUTHORS:20170223-150518455

Funding

NIH
GM61077
NIH
GM120087
NIH
GM65484
NIH
GM118089
NIH Predoctoral Fellowship
T32 GM07616
NIH Predoctoral Fellowship
T32 GM08230
NIH Predoctoral Fellowship
T32 CA009582
NIH
S10 RR025677
Gordon and Betty Moore Foundation

Dates

Created
2017-02-23
Created from EPrint's datestamp field
Updated
2022-04-04
Created from EPrint's last_modified field