We will use computational techniques that were previously developed and tested by our research group in studies of DNA replication and related biochemical processes. These computer simulations will systematically examine changes in the activation barrier of the chemical step due to different factors including the effects of point mutations of distant pol B residues, to substitutions of the p-y bridging oxygen by the - C X Y - groups where X. Y = H, CH3, F, CI, Br or N3, and substitutions in the deoxyribose residues of the primer or dNTP nucleotides. Additionally, calculations of the substrate binding in its ground- and transition-state configurations will be investigated by proven computational strategies with extended simulation times. The results of the simulations (including TS structures and charge distributions as well as simulation protocols) will form a database that will include numerous experimentally-testable predictions along with the list of the assumptions and theoretical models that were used to generate these predictions. This database, which will be made accessible online, will be continually updated with new experimental and computational results, helping in selecting the best models and simulation strategies, and eventually to understanding of the functionally important properties of DNA polymerases at the atomic level. Additionally, the comparative studies will help in selecting the most effective approach for computational inhibitor and mutational screening.

Public Health Relevance

The integration of structural, biochemical and computational studies of DNA replication fidelity requires a collaborative approach, where the theoretical analysis should be treated as an integral part of the experimental effort. Thus, the proposed core will focus on supporting Projects 1-3 with routine calculations of the properties of DNA polymerase B(pol B).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program--Cooperative Agreements (U19)
Project #
1U19CA177547-01
Application #
8591739
Study Section
Special Emphasis Panel (ZCA1-RPRB-B (M2))
Project Start
2013-09-03
Project End
2018-08-31
Budget Start
2013-09-03
Budget End
2014-08-31
Support Year
1
Fiscal Year
2013
Total Cost
$49,721
Indirect Cost
$19,403
Name
University of Southern California
Department
Type
DUNS #
072933393
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
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