Eukaryotic bHLH/PAS (basic helix-loop-helix/Per-ARNT-Sim) transcription factors play a critical role in regulating cellular responses to hypoxia, xenobiotic compounds, and several other environmental conditions. Deregulation of these pathways is highly correlated with several forms of cancer, including solid tumor onset and progression, making their control of particular interest. To this end, the PAS domains within these factors represent particularly attractive targets for small molecule regulation given the role that these domains play as cofactor-regulated protein/protein interaction domains in a wide range of sensory proteins throughout biology. Here we propose the development of an HTS-compatible protein/protein interaction screen to identify compounds that can disrupt the complex between the PAS-B domain of ARNT (aryl hydrocarbon nuclear receptor translocator), a common element of several bHLH/PAS heterodimeric complexes, and coiled-coil segments of several coactivator proteins that are essential for ARNT function. Preliminary low-throughput assays suggest that this approach is feasible for conversion to HTS format, and a combination of in vitro biophysical and cell-based biochemical assays are already in place to validate compounds found in this manner.

Public Health Relevance

We propose to develop an assay for high-throughput screening of small molecule disruptors of interactions between the ARNT/AINT transcriptional activator/coactivator pair. Inappropriate deregulation of this pathway is correlated with a variety of cancers, and the use of this assay to identify artificial compounds which can disrupt this complex will provide novel research tools for the study and manipulation of this complex.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
3R21NS067624-01S1
Application #
8413715
Study Section
Special Emphasis Panel (ZRG1-BST-J (51))
Program Officer
Scheideler, Mark A
Project Start
2009-09-25
Project End
2012-08-31
Budget Start
2009-09-25
Budget End
2012-08-31
Support Year
1
Fiscal Year
2012
Total Cost
$39,625
Indirect Cost
$14,625
Name
University of Texas Sw Medical Center Dallas
Department
Biochemistry
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390