Light-dependent phosphorylation of the drosophila transient receptor potential ion channel.
The Drosophila phototransduction cascade terminates in the opening of an ion channel, designated transient receptor potential (TRP). TRP has been shown to become phosphorylated in vitro, suggesting regulation of the ion channel through posttranslational modification. However, except for one phosphorylation site, Ser(982), which was analyzed by functional in vivo studies (Popescu, D. C., Ham, A. J., and Shieh, B. H. (2006) J. Neurosci. 26, 8570-8577), nothing is known about the role of TRP phosphorylation in vivo. Here, we report the identification of 21 TRP phosphorylation sites by a mass spectrometry approach. 20 phosphorylation sites are located in the C-terminal portion of the channel, and one site is located near the N terminus. All 21 phosphorylation sites were also identified in the inaC(P209) mutant, indicating that phosphorylation of TRP at these sites occurred independently from the eye-enriched protein kinase C. Relative quantification of phosphopeptides revealed that at least seven phosphorylation sites were predominantly phosphorylated in the light, whereas one site, Ser(936), was predominantly phosphorylated in the dark. We show that TRP phosphorylated at Ser(936) was located in the rhabomere. Light-dependent changes in the phosphorylation state of this site occurred within minutes. The dephosphorylation of TRP at Ser(936) required activation of the phototransduction cascade.
Voolstra
Olaf
O
Department of Biosensorics, Institute of Physiology, Germany. voolstra@uni-hohenheim.de
Beck
Katherina
K
Oberegelsbacher
Claudia
C
Pfannstiel
Jens
J
Huber
Armin
A
D000595:Amino Acid Sequence; D000818:Animals; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000528:radiation effects*; D005123:Eye / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D008027:Light; D055537:Light Signal Transduction; D008969:Molecular Sequence Data; D010766:Phosphorylation / Q000528:radiation effects; D010786:Photoreceptor Cells / Q000378:metabolism / Q000528:radiation effects*; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D050051:Transient Receptor Potential Channels / Q000378:metabolism*
D029721:Drosophila Proteins; D050051:Transient Receptor Potential Channels; C063169:trp protein, Drosophila; D011493:Protein Kinase C