The human APOBEC3G (A3G) protein is a DNA deaminase that has potent activity against HIV-1 and a variety of other retroelements. Although HIV encodes a natural inhibitor of A3G called Vif, we hypothesize that the virus still benefits from the mutagenic activity of A3G (immune escape and drug resistance). As an essential step toward testing this hypothesis in vivo, we have adapted, optimized and miniaturized a fluorescence-based DNA cytidine deaminase assay to high throughput format. Preliminary studies have identified 2 candidate inhibitors in the 1280 compound LOPAC library. Here, we propose two specific aims. First, we will collaborate with an NIH Molecular Libraries Probe Production Center in a full high-throughput screen for additional A3G activity inhibitors. Second, we will implement a variety of secondary screens to determine the specificity of the lead actives. We anticipate that the successful completion of the proposed studies will result in the identification and validation of approximately 50 active DNA deaminase inhibitors. A subset of these small molecules will undoubtedly become tools for many future experiments and further development.

Public Health Relevance

The human protein APOBEC3G has become the prototypic DNA cytidine deaminase. The fact that it can mutate HIV cDNA raises the possibility that modulating its activity may have therapeutic merit.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Small Research Grants (R03)
Project #
1R03MH089432-01
Application #
7844122
Study Section
Special Emphasis Panel (ZRG1-BST-J (50))
Program Officer
Yao, Yong
Project Start
2009-09-30
Project End
2011-05-31
Budget Start
2009-09-30
Budget End
2010-05-31
Support Year
1
Fiscal Year
2009
Total Cost
$37,750
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455