The modulation of TRPV4 channel activity through its Ser 824 residue phosphorylation by SGK1

  • Lee, Run-Jeoung (Department of Biology Education, Chungbuk National University) ;
  • Shin, Sung-Hwa (Department of Biology Education, Chungbuk National University) ;
  • Chun, Jae-Sun (Department of Biology Education, Korea National University of Education) ;
  • Hyun, Sung-Hee (Department of Pre-medicine, Eulji University School of Medicine) ;
  • Kim, Yang-Mi (Department of Physiology, College of Medicine, Chungbuk National University) ;
  • Kang, Sang-Sun (Department of Biology Education, Chungbuk National University)
  • Received : 2010.02.17
  • Published : 2010.06.30


With the consensus sequence information of the serum glucocorticoid-induced protein kinase-1 (SGK1) phosphorylation site {R-X-R-X-X-(S/T)$\Phi$; where $\Phi$ is any hydrophobic amino acid}, we noticed that the transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, harbors the putative SGK1 phosphorylation site (on its Ser 824). We have demonstrated that TRPV4 is an SGK1 authentic substrate protein, with the phosphorylation on the Ser 824 of TRPV4 by SGK1. Further, using TRPV4 mutants (S824A and S824D), we noted that the modification of the Ser 824 activates its $Ca^{2+}$ entry, and sensitizes the TRPV4 channel to 4-$\alpha$-phorbol 12,13-didecanoate (4-${\alpha}PDD$) or heat, simultaneously enhancing its active state. Additionally, we determined that the modification of the Ser 824 controls both its plasma membrane localization and its protein interactions with calmodulin. Thus, we have proposed herein that phosphorylation on the Ser 824 of TRPV4 is one of the control points for the regulation of its functions.


Supported by : National Research Foundation of Korea (NRF)


  1. Andrade YN, Fernandes J, Vazquez E, Fernandez-Fernandez JM, Arniges M, Sanchez TM, Villalon M, Valverde MA. 2005. TRPV4 channel is involved in the coupling of fluid viscosity changes to epithelial ciliary activity. J Cell Biol. 168:869-874.
  2. Becker D, Blase C, Bereiter-Hahn J, Jendrach M. 2005. TRPV4 exhibits a functional role in cell-volume regulation. J Cell Sci. 118:2435-2440.
  3. Birder L, Kullmann FA, Lee H, Barrick S, de Groat W, Kanai A, Caterina M. 2007. Activation of urothelial transient receptor potential vanilloid 4 by 4alpha-phorbol 12,13-didecanoate contributes to altered bladder reflexes in the rat. J Pharmacol Exp Ther. 323:227-235.
  4. Cao DS, Yu SQ, Premkumar LS. 2009. Modulation of transient receptor potential Vanilloid 4-mediated membrane currents and synaptic transmission by protein kinase C. Mol Pain. 10:5:1-15.
  5. Casamayor A, Torrance PD, Kobayashi T, Thorner J, Alessi DR. 1999. Functional counterparts of mammalian protein kinases PDK1 and SGK in budding yeast. Curr Biol. 9:186-197.
  6. Chen SY, Bhargava A, Mastroberardino L, Meijer OC, Wang J, Buse P, Firestone GL, Verrey F, Pearce D. 1999. Epithelial sodium channel regulated by aldosterone-induced protein sgk. Proc Natl Acad Sci USA. 96:2514-2519.
  7. Chun J, Kwon T, Kim DJ, Park I, Chung G, Lee EJ, Hong SK, Chang SI, Kim HY, Kang SS. 2003. Inhibition of mitogen-activated kinase kinase kinase 3 activity through phosphorylation by the serum- and glucocorticoid-induced kinase 1. J Biochem. 133:103-108.
  8. Chun J, Kwon T, Lee EJ, Kim CH, Han YS, Hong SK, Hyun S, Kang SS. 2004. 14-3-3 Protein mediates phosphorylation of microtubule-associated protein tau by serum- and glucocorticoid-induced kinase 1. Mol Cells. 18:360-368.
  9. Cohen DM. 2006. Regulation of TRP channels by N-linked glycosylation. Semin Cell Dev Biol. 17:630-637.
  10. Cuajungco MP, Grimm C, Oshima K, D'hoedt D, Nilius B, Mensenkamp AR, Bindels RJ, PI om ann M, Heller S. 2006. PACSINs bind to the TRPV4 cation channel. PACSIN 3 modulates the subcellular localization of TRPV4. J Biol Chem. 281:18753-18762.
  11. Earley S, Heppner TJ, Nelson MT, Brayden JE. 2005. TRPV4 forms a novel $Ca^{2+}$ signaling complex with ryanodine receptors and BKCa channels. Circ Res. 97:1270-1279.
  12. Everaerts W, Nilius B, Owsianik G. 2009. The vallinoid transient receptor potential channel Trpv4: from structure to disease. Prog Biophys Mol Biol. 1:1-25.
  13. Fan HC, Zhang X, McNaughton PA. 2009. Activation of the TRPV4 ion channel is enhanced by phosphorylation. J Biol Chem. 284:27884-27891.
  14. Fernandes J, Lorenzo IM, Andrade YN, Garcia-Elias A, Serra SA, Fernandez-Fernandez JM, Valverde MA. 2008. IP3 sensitizes TRPV4 channel to the mechanoand osmotransducing messenger 5'-6' -epoxyeicosatrienoic acid. J Cell Biol. 181:143-155.
  15. Fu Y, Subramanya A, Rozansky D, Cohen DM. 2006. WNK kinases influence TRPV4 channel function and localization. Am J Physiol Renal Physiol. 290:1305-1314.
  16. Garcia-Elias A, Lorenzo IM, Vicente R, Valverde MA. 2008. $IP_{3}$ receptor binds to and sensitizes TRPV4 channel to osmotic stimuli via a calmodulin-binding site. J Biol Chem. 283:31284-31288.
  17. Hardie RC. 2007. TRP channels and lipids: from Drosophila to mammalian physiology. J Physiol. 578:9-24.
  18. Kobayashi T, Deak M, Morrice N, Cohen P. 1999. Characterization of the structure and regulation of two novel isoforms of serum- and glucocorticoid-induced protein kinase. Biochem J. 1:189-197.
  19. Lambers TT, Weidema AF, Nilius B, Hoenderop JG, Bindels RJ. 2004. Regulation of the mouse epithelial $Ca^{2(+)}$ sensor calmodulin. J Biol Chem. 279:28855-28861.
  20. Lee EJ, Chun J, Hyun S, Ahn HR, Jeong JM, Hong SK, Hong JT, Chang IK, Jeon HY, Han YS, Auh CK, Park JI, Kang SS. 2008. Regulation Fe65 localization to the nucleus by SGK1 phosphorylation of its Ser566 residue. BMB Rep. 41:41-47.
  21. Liedtke W, Choe Y, Marti-Renom MA, Bell AM, Denis CS, Sali A, Hudspeth AJ, Friedman JM, Heller S. 2000. Vanilloid receptor-related osmotically activated channel (VR-OAC), a candidate vertebrate osmoreceptor. Cell. 103:525-535.
  22. Maiyar AC, Huang AJ, Phu PT, Cha HH, Firestone GL. 1996. p53 stimulates promoter activity of the sgk. Serum/glucocorticoid-inucible serin/threonine protein kinase gene in rodent mammary epithelial cells. J Biol Chem. 271:12414-12422.
  23. Murray JT, Cummings LA, Bloomberg GB, Cohen P. 2005. Identification of different specificity requirements between SGK1 and PKBalpha. FEBS Lett. 579:991-994.
  24. Nilius B, Watanabe H, Vriens J. 2003. The TRPV4 channel: structure-function relationship and promiscuous gating behaviour. Pflugers Arch. 446:298-803.
  25. Saito M, Hanson PI, Schlesinger P. 2007. Luminal chloride-dependent activation of endosome calcium channels: patch clamp study of enlarged endosomes. J Biol Chem. 282:27327-27333.
  26. Strotmann R, Schultz G, Plant TD. 2003. Ca-dependent potentiation of thenonselective cation channel TRPV4 is mediated by a Csterminal calmodulin binding site. J Biol Chem. 278:26541-26549.
  27. Suzuki M, Hirao A, Mizuno A. 2003a. Microtubule-associated [corrected] protein 7 increase the membrane expression of transient receptor potential vanilloid 4 (TRPV4). J Biol Chem. 278:51448-51453.
  28. Suzuki M, Mizuno A, Kodaira K, Imai M. 2003b. Impaired pressure sensation in mice lacking TRPV4. J Biol Chem. 278:22664-22668.
  29. Tessier M, Woodgett JR. 2006. Serum and Glucocorticoid-regulated protein kinases: variations on a theme. J Cell Biochem. 98:1391-1407.
  30. van de Graaf SF, Hoenderop JG, Bindels RJ. 2006. Regulation TRPV5 and TRPV6 by associated proteins. Am J Physiol Renal Physiol. 290:1295-1302.
  31. Vennekens R, Owsianik G, Nilius B. 2008. Vanilloid transient receptor potential cation channels: an overview. Curr Pharm Des. 14:18-31.
  32. Vriens J, Janssens A, Prenen J, Nilius B, Wondergem R. 2006. TRPV channels and modulation by hepatocyte growth factor/scatter factor in human hepatoblastoma (HepG2) cells. Cell Calcium. 36:19-28.
  33. Vriens J, Owsianik G, Janssens A, Voets T, Nilius B. ?2007. Determinants of 4 alpha-phorbol sensitivity in transmembrane domains 3 and 4 of the cation channel TRPV4. J Biol Chem. 282:12796-12803.
  34. Watanabe H, Davis JB, Smart D, Jerman JC, Smith GD, Hayes P, Vriens J, Cairns W, Wissenbach U, Prenen J, Flockerzi V, Droogmans G, Benham CD, Nilius B. 2002. Activation of TRPV4 channels (hVRL-2/mTRPl2) by phorbol derivatives. J Biol Chem. 277:13569-13577.
  35. Watanabe H, Vriens J, Prenen J, Droogmans G, Voets T, Nilius B. 2003. Anandamide and arachidonic acid use epoxyeicosatrienoic acids to activate TRPV 4 channels. Nature. 424:434-438.
  36. Webster MK, Goya L, Ge Y, Maiyar AC, Firestone GL. 1993. Characterization of sgk, a novel member of the serine/threonine protein kinase gene family which is transcriptionally induced by glucocorticoids and serum. Mol Cell Biol. 13:2031-2040.
  37. Wegierski T, Lewandrowski U, Muller B, Sickmann A, Walz G. 2009. Tyrosin phosphorylation modulates the activity of TRPV4 in response to defined stimuli. J Biol Chem. 284:2923-2933.
  38. Xu F, Satoh E, Iijima T. 2003. Protein kinase C-mediated $Ca^{2+}$ entry in HEK 293cells transiently expressing human TRPV4. Br J Pharmacol. 140:413-421.
  39. Yoshida T, Inoue R, Morii T, Takahashi N, Yamamoto S, Hara Y, Tominaga M, Shimizu S, Sato Y, Mori Y. 2006. Nitric oxide activates TRP channels by cysteine S-nitryosylation. Nat Chern Biol. 2:596-607.