The beta 1 and beta 2 adrenergic receptors are members of the super-family of G protein-coupled receptors (GPCRs). This receptor super-family comprises some of the most important pharmacological targets for the treatment of cardiovascular (e.g. heart failure, hypertension) and pulmonary diseases (e.g. asthma). A major cause for the observed attenuation of adrenergic response in heart failure and asthma is receptor downregulation, which results from chronic agonist exposure. Recently, ubiquitination of cell-surface receptors has been implicated as an important mechanism for post-endocytic sorting to lysosomes. Ubiquitination is a post-translational modification of proteins orchestrated by a well-defined process involving a cascade of three enzymatic activities. Of these the final step, catalyzed by the enzyme, E3 ubiquitin ligase, determines the specificity of substrate ubiquitination. We have demonstrated that two representative GPCRs, the beta2 adrenergic receptor and the V2 vasopressin receptor and their adaptor protein beta-arrestin become ubiquitinated upon agonist stimulation. Ubiquitination of beta-arrestin is crucial for receptor internalization, whereas ubiquitination of the receptor is essential for the proper sorting and downregulation of the internalized and activated receptors. We hypothesize that ubiquitination and deubiquitination coordinate the trafficking and signaling of GPCRs and involve a specific set of endocytic adapter proteins including beta-arrestins.
The specific aims are: 1) to define the molecular mechanisms of receptor ubiquitination leading to receptor downregulation, and 2) to determine the roles of deubiquitinating enzyme(s) in receptor trafficking. Unraveling the molecular mechanisms governing the regulation of GPCRs, especially beta ARs, by ubiquitination could have a great impact on the development of novel therapeutic strategies for cardiovascular and pulmonary diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL080525-03
Application #
7269341
Study Section
Hypertension and Microcirculation Study Section (HM)
Program Officer
Buxton, Denis B
Project Start
2005-08-01
Project End
2010-07-31
Budget Start
2007-08-01
Budget End
2008-07-31
Support Year
3
Fiscal Year
2007
Total Cost
$258,854
Indirect Cost
Name
Duke University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Jean-Charles, Pierre-Yves; Kaur, Suneet; Shenoy, Sudha K (2017) G Protein-Coupled Receptor Signaling Through ?-Arrestin-Dependent Mechanisms. J Cardiovasc Pharmacol 70:142-158
Jean-Charles, Pierre-Yves; Yu, Samuel Mon-Wei; Abraham, Dennis et al. (2017) Mdm2 regulates cardiac contractility by inhibiting GRK2-mediated desensitization of ?-adrenergic receptor signaling. JCI Insight 2:
Jean-Charles, P-Y; Snyder, J C; Shenoy, S K (2016) Chapter One - Ubiquitination and Deubiquitination of G Protein-Coupled Receptors. Prog Mol Biol Transl Sci 141:1-55
Jean-Charles, P-Y; Freedman, N J; Shenoy, S K (2016) Chapter Nine - Cellular Roles of Beta-Arrestins as Substrates and Adaptors of Ubiquitination and Deubiquitination. Prog Mol Biol Transl Sci 141:339-69
Shenoy, Sudha K (2016) Preface. Prog Mol Biol Transl Sci 141:xiii-xiv
Jean-Charles, Pierre-Yves; Rajiv, Vishwaesh; Shenoy, Sudha K (2016) Ubiquitin-Related Roles of ?-Arrestins in Endocytic Trafficking and Signal Transduction. J Cell Physiol 231:2071-80
Feger, Bryan J; Thompson, J Will; Dubois, Laura G et al. (2016) Microgravity induces proteomics changes involved in endoplasmic reticulum stress and mitochondrial protection. Sci Rep 6:34091
Jean-Charles, Pierre-Yves; Rajiv, Vishwaesh; Shenoy, Sudha K (2016) Cover Image, Volume 231, Number 10, October 2016. J Cell Physiol 231:i
Jean-Charles, Pierre-Yves; Zhang, Lisheng; Wu, Jiao-Hui et al. (2016) Ubiquitin-specific Protease 20 Regulates the Reciprocal Functions of ?-Arrestin2 in Toll-like Receptor 4-promoted Nuclear Factor ?B (NF?B) Activation. J Biol Chem 291:7450-64
Kommaddi, Reddy Peera; Jean-Charles, Pierre-Yves; Shenoy, Sudha K (2015) Phosphorylation of the deubiquitinase USP20 by protein kinase A regulates post-endocytic trafficking of ?2 adrenergic receptors to autophagosomes during physiological stress. J Biol Chem 290:8888-903

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