The conformational dynamics of transcriptional activators, coactivators and their complexes underpin regulated transcription. The low energy barriers between individual conformations mean that each participant can use the same group of amino acids to recognize a variety of binding partners, with each complex assuming a distinct conformation. The assumption is that each of the conformations correlates with a unique functional outcome via allosteric communication with other binding partners. However, this model has never been tested, primarily due to a lack of robust tools to connect in vitro observations with function in cells. Our goal is to develop and implement chemical genetic tools for this purpose. Focusing on two functionally important coactivator motifs, KIX (neuropathic pain, neurodegenerative disorders) and AcID (cancer, viral infection), we will develop covalent chemical co-chaperones that stabilize particular conformations of the coactivators alone and in complex with cognate ligands. These co-chaperones will be used to rigorously characterize the structure (X-ray crystallography, NMR spectroscopy) and dynamics (transient kinetics, equilibrium binding, computational analysis) of the coactivators in distinct conformational and assembly states. Complementing the in vitro data will be experiments with chemical co-chaperones in cells, thus identifying both the interactions and the conformations that regulate transcriptional output. Through these studies, allosteric binding sites for small molecule targeting of these disease-relevant transcriptional complexes will be identified.

Public Health Relevance

Transcription is dysregulated in every human disease as either a cause or an effect and as such represents a potentially powerful intervention point for therapeutic development. Here we will develop chemical co?chaperones that target individual transcriptional coactivators and use those tools to define the interactions and conformational changes that produce function in vitro and in cells. In doing so, we will discovery new targets for drug discovery, with a particular focus on transcriptional pathways integral to neuropathic pain and cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM065330-10
Application #
8964620
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Fabian, Miles
Project Start
2002-04-01
Project End
2019-02-28
Budget Start
2015-07-01
Budget End
2016-02-29
Support Year
10
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
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