The decrease of cell responsiveness to a persistent stimulus, usually termed desensitization, is a widespread biological phenomenon. Visual amplification cascade (and the signaling by other G protein-coupled receptors) is attenuated by a two-step mechanism: phosphorylation of light-activated rhodopsin (Rh*) by rhodopsin kinase followed by arrestin binding to light-activated phosphorylated rhodopsin (P-Rh*). Arrestin binding terminates transducin-mediated signaling, playing an important role in the recovery of photoreceptor cells. The main objective of this proposal is to elucidate how the fine molecular mechanisms of visual arrestin function translate into its timely binding to rhodopsin, subsequent dissociation from phosphoopsin, its translocation into rod outer segment in the light and its movement to the inner segment in the dark. Using site-directed mutagenesis, direct binding assay, spin labeling of arrestin and rhodopsin followed by EPR spectroscopy, and X-ray crystallography we will identify arrestin and rhodopsin residues participating in their interaction. We will elucidate the number of rhodopsin-attached phosphates necessary for tight arrestin binding and the role of arrestin and rhodopsin dimerization in their interaction. We will use custom-designed arrestin mutants expressed in transgenic mice to study the kinetics of signal shut-off and recovery in rods, the physiological role of arrestin self-association and function of its light-dependent translocation. Several congenital disorders are associated with excessive rhodopsin signaling. We have created phosphorylation-independent """"""""super-arrestins"""""""" binding with high affinity to P-Rh* and Rh*, that appear to be logical tools for gene therapy of these disorders. The feasibility of using """"""""super-arrestins"""""""" as tools for correcting response kinetics and preventing retinal degeneration in mouse models with the loss of rhodopsin phosphorylation sites or rhodopsin kinase will be also tested.
Tso, Shih-Chia; Chen, Qiuyan; Vishnivetskiy, Sergey A et al. (2018) Using two-site binding models to analyze microscale thermophoresis data. Anal Biochem 540-541:64-75 |
Vishnivetskiy, Sergey A; Sullivan, Lori S; Bowne, Sara J et al. (2018) Molecular Defects of the Disease-Causing Human Arrestin-1 C147F Mutant. Invest Ophthalmol Vis Sci 59:13-20 |
Gurevich, Vsevolod V; Gurevich, Eugenia V (2018) GPCRs and Signal Transducers: Interaction Stoichiometry. Trends Pharmacol Sci 39:672-684 |
Cleghorn, Whitney M; Bulus, Nada; Kook, Seunghyi et al. (2018) Non-visual arrestins regulate the focal adhesion formation via small GTPases RhoA and Rac1 independently of GPCRs. Cell Signal 42:259-269 |
Gurevich, Vsevolod V; Gurevich, Eugenia V; Uversky, Vladimir N (2018) Arrestins: structural disorder creates rich functionality. Protein Cell 9:986-1003 |
Chen, Qiuyan; Iverson, Tina M; Gurevich, Vsevolod V (2018) Structural Basis of Arrestin-Dependent Signal Transduction. Trends Biochem Sci 43:412-423 |
Vishnivetskiy, Sergey A; Lee, Regina J; Zhou, X Edward et al. (2017) Functional role of the three conserved cysteines in the N domain of visual arrestin-1. J Biol Chem 292:12496-12502 |
Sullivan, Lori S; Bowne, Sara J; Koboldt, Daniel C et al. (2017) A Novel Dominant Mutation in SAG, the Arrestin-1 Gene, Is a Common Cause of Retinitis Pigmentosa in Hispanic Families in the Southwestern United States. Invest Ophthalmol Vis Sci 58:2774-2784 |
Zhu, Lu; AlmaƧa, Joana; Dadi, Prasanna K et al. (2017) ?-arrestin-2 is an essential regulator of pancreatic ?-cell function under physiological and pathophysiological conditions. Nat Commun 8:14295 |
Zhou, X Edward; He, Yuanzheng; de Waal, Parker W et al. (2017) Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors. Cell 170:457-469.e13 |
Showing the most recent 10 out of 144 publications