Phosphorylation of rhodopsin critically controls the visual transduction cascade by uncoupling it from the G-protein transducin. The kinase primarily responsible for this phosphorylation is rhodopsin kinase, a substrate-regulated kinase that phosphorylates light-activated rhodopsin. Protein kinase C has been implicated in controlling the phosphorylation of both light-activated and dark-adapted rhodopsin. Two of the major rhodopsin phosphorylation sites in vivo, Ser334 and Ser338, are effective protein kinase C phosphorylation sites in vitro, while the latter is preferentially phosphorylated by rhodopsin kinase in vitro. Using phosphospecific antibodies against each of these two sites, we show that both sites are under differential spatial and temporal regulation. Exposure of mice to light results in rapid phosphorylation of Ser338 that is evenly distributed along the rod outer segment. Phosphorylation of Ser334 is considerably slower, begins at the base of the rod outer segment, and spreads to the top of the photoreceptor over time. In addition, we show that phosphorylation of both sites is abolished in rhodopsin kinase−/− mice, revealing an absolute requirement for rhodopsin kinase to phosphorylate rhodopsin. This requirement may reflect the need for priming phosphorylations at rhodopsin kinase sites allowing for subsequent phosphorylation by protein kinase C at Ser334. In this regard, treatment of mouse retinas with phorbol esters results in a 4-fold increase in phosphorylation on Ser334, with no significant effect on the phosphorylation of Ser338. Our results are consistent with light triggering rapid priming phosphorylations of rhodopsin by rhodopsin kinase, followed by a slower phosphorylation on Ser334, which is regulated by protein kinase C.
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September 2003
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Research Article|
September 01 2003
Differential spatial and temporal phosphorylation of the visual receptor, rhodopsin, at two primary phosphorylation sites in mice exposed to light
Ryan A. ADAMS;
Ryan A. ADAMS
∗Department of Chemistry and Biochemistry, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0640, U.S.A.
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Xinran LIU;
Xinran LIU
†Department of Pharmacology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0640, USA
‡Center for Basic Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9111, U.S.A.
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David S. WILLIAMS;
David S. WILLIAMS
†Department of Pharmacology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0640, USA
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Alexandra C. NEWTON
Alexandra C. NEWTON
1
†Department of Pharmacology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0640, USA
1To whom correspondence should be addressed (e-mail anewton@ucsd.edu).
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Publisher: Portland Press Ltd
Received:
March 13 2003
Revision Received:
June 05 2003
Accepted:
June 16 2003
Accepted Manuscript online:
June 16 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 374 (2): 537–543.
Article history
Received:
March 13 2003
Revision Received:
June 05 2003
Accepted:
June 16 2003
Accepted Manuscript online:
June 16 2003
Citation
Ryan A. ADAMS, Xinran LIU, David S. WILLIAMS, Alexandra C. NEWTON; Differential spatial and temporal phosphorylation of the visual receptor, rhodopsin, at two primary phosphorylation sites in mice exposed to light. Biochem J 1 September 2003; 374 (2): 537–543. doi: https://doi.org/10.1042/bj20030408
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