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Phosphorylation of the Regulator of G Protein Signaling RGS9-1 by Protein Kinase A Is a Potential Mechanism of Light- and Ca2+-Mediated Regulation of G Protein Function in Photoreceptors

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Department of Molecular and Cellular Pharmacology and Neuroscience Program, University of Miami School of Medicine, Miami, Florida 33136, and Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, UMR 6097, Valbonne, France
Cite this: Biochemistry 2001, 40, 42, 12619–12627
Publication Date (Web):September 26, 2001
Copyright © 2001 American Chemical Society
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In vertebrate photoreceptors, photoexcited rhodopsin interacts with the G protein transducin, causing it to bind GTP and stimulate the enzyme cGMP phosphodiesterase. The rapid termination of the active state of this pathway is dependent upon a photoreceptor-specific regulator of G protein signaling RGS9-1 that serves as a GTPase activating protein (GAP) for transducin. Here, we show that, in preparations of photoreceptor outer segments (OS), RGS9-1 is readily phosphorylated by an endogenous Ser/Thr protein kinase. Protein kinase C and MAP kinase inhibitors reduced labeling by about 30%, while CDK5 and CaMK II inhibitors had no effect. cAMP-dependent protein kinase (PKA) inhibitor H89 reduced RGS9-1 labeling by more than 90%, while dibutyryl-cAMP stimulated it 3-fold, implicating PKA as the major kinase responsible for RGS9-1 phosphorylation in OS. RGS9-1 belongs to an RGS subfamily also including RGS6, RGS7, and RGS11, which exist as heterodimers with the G protein β subunit Gβ5. Phosphorylated RGS9-1 remains associated with Gβ5L, a photoreceptor-specific splice form, which itself was not phosphorylated. RGS9-1 immunoprecipitated from OS was in vitro phosphorylated by exogenous PKA. The PKA catalytic subunit could also phosphorylate recombinant RGS9-1, and mutational analysis localized phosphorylation sites to Ser427 and Ser428. Substitution of these residues for Glu, to mimic phosphorylation, resulted in a reduction of the GAP activity of RGS9-1. In OS, RGS9-1 phosphorylation required the presence of free Ca2+ ions and was inhibited by light, suggesting that RGS9-1 phosphorylation could be one of the mechanisms mediating a stronger photoresponse in dark-adapted cells.

 Supported by NIH RO1 Grants GM60019 and EY12982 and Fight for Sight Research to Prevent Blindness America Foundation (to V.Z.S.) and by a Young Investigator Award from the National Alliance for Research on Schizophrenia and Depression (NARSAD) (to K.L.)

 Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine.


 Institut de Pharmacologie Moleculaire et Cellulaire, CNRS.


 Address correspondence to this author at the University of Miami School of Medicine, R-189, 1600 NW 10th Ave., Miami, FL 33136. Phone:  (305) 243-3430. Fax:  (305) 243-4555. E-mail:  [email protected]

 Neuroscience Program, University of Miami School of Medicine.

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  1. Shinya Sato, Takahiro Yamashita, Michiyuki Matsuda. Rhodopsin-mediated light-off-induced protein kinase A activation in mouse rod photoreceptor cells. Proceedings of the National Academy of Sciences 2020, 117 (43) , 26996-27003.
  2. Gabriel Peinado Allina, Christopher Fortenbach, Franklin Naarendorp, Owen P. Gross, Edward N. Pugh, Marie E. Burns. Bright flash response recovery of mammalian rods in vivo is rate limited by RGS9. Journal of General Physiology 2017, 149 (4) , 443-454.
  3. Ching-Kang Chen, Michael L. Woodruff, Gordon L. Fain. Rhodopsin kinase and recoverin modulate phosphodiesterase during mouse photoreceptor light adaptation. Journal of General Physiology 2015, 145 (3) , 213-224.
  4. Ching-Kang Jason Chen. RGS Protein Regulation of Phototransduction. 2015,,, 31-45.
  5. Mei Tian, Marisa Zallocchi, Weimin Wang, Ching-Kang Chen, Krzysztof Palczewski, Duane Delimont, Dominic Cosgrove, You-Wei Peng, . Light-Induced Translocation of RGS9-1 and Gβ5L in Mouse Rod Photoreceptors. PLoS ONE 2013, 8 (3) , e58832.
  6. Jacob Kach, Nan Sethakorn, Nickolai O. Dulin. A finer tuning of G-protein signaling through regulated control of RGS proteins. American Journal of Physiology-Heart and Circulatory Physiology 2012, 303 (1) , H19-H35.
  7. V. I. Govardovskii, M. L. Firsov. Unknown Mechanisms Regulating the GPCR Signal Cascade in Vertebrate Photoreceptors. Neuroscience and Behavioral Physiology 2012, 42 (2) , 180-192.
  8. Christina Kiel, Andreas Vogt, Anne Campagna, Andrew Chatr‐aryamontri, Magdalena Swiatek‐de Lange, Monika Beer, Sylvia Bolz, Andreas F Mack, Norbert Kinkl, Gianni Cesareni, Luis Serrano, Marius Ueffing. Structural and functional protein network analyses predict novel signaling functions for rhodopsin. Molecular Systems Biology 2011, 7 (1) , 551.
  9. Z. Xie, Z. Yang, K. M. Druey. Phosphorylation of RGS13 by the Cyclic AMP-dependent Protein Kinase Inhibits RGS13 Degradation. Journal of Molecular Cell Biology 2010, 2 (6) , 357-365.
  10. Norma M. Giusto, Susana J. Pasquaré, Gabriela A. Salvador, Mónica G. Ilincheta de Boschero. Lipid second messengers and related enzymes in vertebrate rod outer segments. Journal of Lipid Research 2010, 51 (4) , 685-700.
  11. John Traynor. Regulator of G protein-signaling proteins and addictive drugs. Annals of the New York Academy of Sciences 2010, 1187 (1) , 341-352.
  12. Anja Ruppelt, Nikolaus G. Oberprieler, George Magklaras, Kjetil Taskén. Physiological Substrates of PKA and PKG. 2010,,, 1497-1514.
  13. Xiangmin Peng, Zhigang Zhou, Jian Hu, David J. Fink, Marina Mata. Soluble Nogo Receptor Down-regulates Expression of Neuronal Nogo-A to Enhance Axonal Regeneration. Journal of Biological Chemistry 2010, 285 (4) , 2783-2795.
  14. Vladlen Z. Slepak. Chapter 6 Structure, Function, and Localization of Gβ5–RGS Complexes. 2009,,, 157-203.
  15. Luba A. Astakhova, Michael L. Firsov, Victor I. Govardovskii. Kinetics of Turn-offs of Frog Rod Phototransduction Cascade. Journal of General Physiology 2008, 132 (5) , 587-604.
  16. Jing Qiao, Oksana Holian, Bao-Shiang Lee, Fei Huang, Jihang Zhang, Hazel Lum. Phosphorylation of GTP dissociation inhibitor by PKA negatively regulates RhoA. American Journal of Physiology-Cell Physiology 2008, 295 (5) , C1161-C1168.
  17. Elliott M. Ross. Coordinating Speed and Amplitude in G-Protein Signaling. Current Biology 2008, 18 (17) , R777-R783.
  18. Benjamin P.C. Allen, Paul H.J. Nederkoorn, Henk Timmerman, David Timms, Kenneth J. Broadley, Robin H. Davies. Monocation-driven proton transfer relays within G protein-coupled receptors of the rhodopsin class and the GTP synthase mechanism. Journal of Molecular Structure: THEOCHEM 2008, 859 (1-3) , 51-68.
  19. Alecia K. Gross, Qiong Wang, Theodore G. Wensel. Regulation of Photoresponses by Phosphorylation. 2008,,, 125-140.
  20. Patrick Osei-Owusu, Xiaoguang Sun, Ryan M. Drenan, Thomas H. Steinberg, Kendall J. Blumer. Regulation of RGS2 and Second Messenger Signaling in Vascular Smooth Muscle Cells by cGMP-dependent Protein Kinase. Journal of Biological Chemistry 2007, 282 (43) , 31656-31665.
  21. Seena K. Ajit, Suneela Ramineni, Wade Edris, Rachel A. Hunt, Wah-Tung Hum, John R. Hepler, Kathleen H. Young. RGSZ1 interacts with protein kinase C interacting protein PKCI-1 and modulates mu opioid receptor signaling. Cellular Signalling 2007, 19 (4) , 723-730.
  22. Gabriela A. Salvador, Norma M. Giusto. Phospholipase D from photoreceptor rod outer segments is a downstream effector of RhoA: Evidence of a light-dependent mechanism. Experimental Eye Research 2006, 83 (1) , 202-211.
  23. Gary B. Willars. Mammalian RGS proteins: Multifunctional regulators of cellular signalling. Seminars in Cell & Developmental Biology 2006, 17 (3) , 363-376.
  24. Thierry J. Horner, Shoji Osawa, Michael D. Schaller, Ellen R. Weiss. Phosphorylation of GRK1 and GRK7 by cAMP-dependent Protein Kinase Attenuates Their Enzymatic Activities. Journal of Biological Chemistry 2005, 280 (31) , 28241-28250.
  25. Young Hee Kim, Yoon Sook Kim, Hae Sook Noh, Sang Soo Kang, Eun Woo Cheon, Sang Kyu Park, Byung Ju Lee, Wan Sung Choi, Gyeong Jae Cho. Changes in rhodopsin kinase and transducin in the rat retina in early-stage diabetes. Experimental Eye Research 2005, 80 (6) , 753-760.
  26. Evan L. Riddle, Raúl A. Schwartzman, Meredith Bond, Paul A. Insel. Multi-Tasking RGS Proteins in the Heart. Circulation Research 2005, 96 (4) , 401-411.
  27. Javier Garzón, María Rodríguez-Muñoz, Almudena López-Fando, Pilar Sánchez-Blázquez. Activation of μ-Opioid Receptors Transfers Control of Gα Subunits to the Regulator of G-protein Signaling RGS9-2. Journal of Biological Chemistry 2005, 280 (10) , 8951-8960.
  28. C. K. Chen. The vertebrate phototransduction cascade: amplification and termination mechanisms. 2005,,, 101-121.
  29. Guang Hu, Zhixian Zhang, Theodore G. Wensel. Activation of RGS9-1GTPase Acceleration by Its Membrane Anchor, R9AP. Journal of Biological Chemistry 2003, 278 (16) , 14550-14554.
  30. Izabela Sokal, Guang Hu, Yan Liang, Muling Mao, Theodore G. Wensel, Krzysztof Palczewski. Identification of Protein Kinase C Isozymes Responsible for the Phosphorylation of Photoreceptor-specific RGS9-1 at Ser475. Journal of Biological Chemistry 2003, 278 (10) , 8316-8325.
  31. Thomas Wieland, Clemens Mittmann. Regulators of G-protein signalling: multifunctional proteins with impact on signalling in the cardiovascular system. Pharmacology & Therapeutics 2003, 97 (2) , 95-115.
  32. Kjetil Taskén, Anja Ruppelt, Cathrine R. Carlson, John Shabb. Physiological Substrates of PKA and PKG. 2003,,, 501-510.
  33. D. Scott Witherow, Vladlen Z. Slepak. A Novel Kind of G Protein Heterodimer: The Gβ5-RGS Complex. Receptors and Channels 2003, 9 (3) , 205-212.
  34. Peter Chidiac, Anju A. Roy. Activity, Regulation, and Intracellular Localization of RGS Proteins. Receptors and Channels 2003, 9 (3) , 135-147.
  35. K.Saidas Nair, Nagaraj Balasubramanian, Vladlen Z. Slepak. Signal-Dependent Translocation of Transducin, RGS9-1-Gβ5L Complex, and Arrestin to Detergent-Resistant Membrane Rafts in Photoreceptors. Current Biology 2002, 12 (5) , 421-425.
  36. James B. Hurley. Shedding Light on Adaptation. Journal of General Physiology 2002, 119 (2) , 125-128.

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