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Investigation of the role of Mg2+ in DNA repair proteins APE1, Pol ?, and FEN1


EMSL Project ID
10600c

Abstract

We propose to utilize unique capabilities within the EMSL (low temperature solid-state NMR and the mpp2 super computer) to investigate Mg2+ binding to two DNA repair proteins Pol ? and APE1. In the case of Pol ? we are interested in the order of binding of Mg2+ with respect to the binding of damaged DNA. The binding of Mg2+ to Pol ? in the absence of DNA could explain the high error rate of this polymerase. In the case of APE1, the Mg2+ NMR experiments will resolve a key mechanistic question relative to APE1, i.e. does it operate via a single- or two-metal mechanism. To perform and analyze the results of these experiments we need access to 12 weeks (2016 hours) of wide bore 500 time and a similar amount of time on the 400 wide bore spectrometer. For analysis of the results we need 75,000 cpu-hours of mpp2 time.

Project Details

Project type
Capability Research
Start Date
2006-04-06
End Date
2007-05-21
Status
Closed

Team

Principal Investigator

Paul Ellis
Institution
Pacific Northwest National Laboratory

Team Members

Gerard Harbison
Institution
University of Nebraska - Lincoln

Robert Heck
Institution
Pacific Northwest National Laboratory

Samuel Wilson
Institution
National Institute of Environmental Health Sciences, NIH

David Wilson
Institution
Hasselt University

Andrew Lipton
Institution
Environmental Molecular Sciences Laboratory

Related Publications

Lipton AS, RW Heck, SV Primak, DR McNeill, DM Wilson Iii, and PD Ellis. 2008. "Characterization of Mg2+ Binding to the DNA Repair Protein Apurinic/Apyrimidic Endonuclease 1 via Solid-State 25Mg NMR Spectroscopy." Journal of the American Chemical Society 130(29):9332-9341. doi:10.1021/ja0776881