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Light-dependent phosphorylation of the carboxy tail of mouse melanopsin

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Abstract

Melanopsin-based phototransduction is involved in non-image forming light responses including circadian entrainment, pupil constriction, suppression of pineal melatonin synthesis, and direct photic regulation of sleep in vertebrates. Given that the functions of melanopsin involve the measurement and summation of total environmental luminance, there would appear to be no need for the rapid deactivation typical of other G-protein coupled receptors. In this study, however, we demonstrate that heterologously expressed mouse melanopsin is phosphorylated in a light-dependent manner, and that this phosphorylation is involved in regulating the rate of G-protein activation and the lifetime of melanopsin’s active state. Furthermore, we provide evidence for light-dependent phosphorylation of melanopsin in the mouse retina using an in situ proximity ligation assay. Finally, we demonstrate that melanopsin preferentially interacts with the GRK2/3 family of G-protein coupled receptor kinases through co-immunoprecipitation assays. Based on the complement of G-protein receptor kinases present in the melanopsin-expressing retinal ganglion cells, GRK2 emerges as the best candidate for melanopsin’s cognate GRK.

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Abbreviations

GPCR:

G-protein coupled receptor

GRK:

G-protein coupled receptor kinase

PLA:

Proximity-dependent ligation assay

ipRGC:

Intrinsically photosensitive retinal ganglion cell

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Acknowledgments

This study was supported by Grants from the National Science Foundation to P.R.R (IOS0721608), and from the National Eye Institute to P. R. R. (R01EY019053). J. B. was supported by an NIH training grant (T32 GM066706). We would like to thank R. Crouch for the gift of 11-cis-retinal and D. Ujla for constructing the phospho-null melanopsin mutant.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Maryland, Baltimore County, Baltimore, MD, 21250, USA

    Joseph R. Blasic Jr. & Phyllis R. Robinson

  2. Department of Veterinary and Comparative Anatomy, Pharmacology, and Physiology, Washington State University, Pullman, WA, 99164, USA

    R. Lane Brown

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  1. Joseph R. Blasic Jr.
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Correspondence to Phyllis R. Robinson.

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Blasic, J.R., Lane Brown, R. & Robinson, P.R. Light-dependent phosphorylation of the carboxy tail of mouse melanopsin. Cell. Mol. Life Sci. 69, 1551–1562 (2012). https://doi.org/10.1007/s00018-011-0891-3

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