The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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$Ca^{2+}$ is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line

  • Park, Soonhong (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry) ;
  • Ku, Sang Kyun (Department of Oral Medicine, Yonsei University College of Dentistry) ;
  • Ji, Hye Won (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry) ;
  • Choi, Jong-Hoon (Department of Oral Medicine, Yonsei University College of Dentistry) ;
  • Shin, Dong Min (Department of Oral Biology, BK21 PLUS Project, Yonsei University College of Dentistry)
  • Received : 2014.12.23
  • Accepted : 2015.02.25
  • Published : 2015.05.01
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Abstract

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that $Ca^{2+}$ may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced $[Ca^{2+}]_i$ increase with nonspecific $Ca^{2+}$ channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive $[Ca^{2+}]_i$ elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a $Ca^{2+}$ chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that $Ca^{2+}$ may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream $Ca^{2+}$ regulation of the WNK-OSR1 pathway in intact cells.

Keywords

References

  1. Xu B, English JM, Wilsbacher JL, Stippec S, Goldsmith EJ, Cobb MH. WNK1, a novel mammalian serine/threonine protein kinase lacking the catalytic lysine in subdomain II. J Biol Chem. 2000;275:16795-16801. https://doi.org/10.1074/jbc.275.22.16795
  2. Verissimo F, Jordan P. WNK kinases, a novel protein kinase subfamily in multi-cellular organisms. Oncogene. 2001;20:5562-5569. https://doi.org/10.1038/sj.onc.1204726
  3. Wilson FH, Disse-Nicodeme S, Choate KA, Ishikawa K, Nelson-Williams C, Desitter I, Gunel M, Milford DV, Lipkin GW, Achard JM, Feely MP, Dussol B, Berland Y, Unwin RJ, Mayan H, Simon DB, Farfel Z, Jeunemaitre X, Lifton RP. Human hypertension caused by mutations in WNK kinases. Science. 2001;293:1107-1112. https://doi.org/10.1126/science.1062844
  4. Gamba G. Role of WNK kinases in regulating tubular salt and potassium transport and in the development of hypertension. Am J Physiol Renal Physiol. 2005;288:F245-252. https://doi.org/10.1152/ajpcell.00411.2004
  5. Kahle KT, Wilson FH, Leng Q, Lalioti MD, O'Connell AD, Dong K, Rapson AK, MacGregor GG, Giebisch G, Hebert SC, Lifton RP. WNK4 regulates the balance between renal NaCl reabsorption and $K^+$ secretion. Nat Genet. 2003;35:372-376. https://doi.org/10.1038/ng1271
  6. Vitari AC, Deak M, Morrice NA, Alessi DR. The WNK1 and WNK4 protein kinases that are mutated in Gordon's hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. Biochem J. 2005;391:17-24. https://doi.org/10.1042/BJ20051180
  7. Rinehart J, Kahle KT, de Los Heros P, Vazquez N, Meade P, Wilson FH, Hebert SC, Gimenez I, Gamba G, Lifton RP. WNK3 kinase is a positive regulator of NKCC2 and NCC, renal cation-Clcotransporters required for normal blood pressure homeostasis. Proc Natl Acad Sci U S A. 2005;102:16777-16782. https://doi.org/10.1073/pnas.0508303102
  8. Moriguchi T, Urushiyama S, Hisamoto N, Iemura S, Uchida S, Natsume T, Matsumoto K, Shibuya H. WNK1 regulates phosphorylation of cation-chloride-coupled cotransporters via the STE20-related kinases, SPAK and OSR1. J Biol Chem. 2005;280:42685-42693. https://doi.org/10.1074/jbc.M510042200
  9. Kahle KT, Macgregor GG, Wilson FH, Van Hoek AN, Brown D, Ardito T, Kashgarian M, Giebisch G, Hebert SC, Boulpaep EL, Lifton RP. Paracellular Cl- permeability is regulated by WNK4 kinase: insight into normal physiology and hypertension. Proc Natl Acad Sci U S A. 2004;101:14877-14882. https://doi.org/10.1073/pnas.0406172101
  10. Yamauchi K, Rai T, Kobayashi K, Sohara E, Suzuki T, Itoh T, Suda S, Hayama A, Sasaki S, Uchida S. Disease-causing mutant WNK4 increases paracellular chloride permeability and phosphorylates claudins. Proc Natl Acad Sci U S A. 2004;101: 4690-4694. https://doi.org/10.1073/pnas.0306924101
  11. Xu BE, Min X, Stippec S, Lee BH, Goldsmith EJ, Cobb MH. Regulation of WNK1 by an autoinhibitory domain and autophosphorylation. J Biol Chem. 2002;277:48456-48462. https://doi.org/10.1074/jbc.M207917200
  12. Lenertz LY, Lee BH, Min X, Xu BE, Wedin K, Earnest S, Goldsmith EJ, Cobb MH. Properties of WNK1 and implications for other family members. J Biol Chem. 2005;280:26653-26658. https://doi.org/10.1074/jbc.M502598200
  13. Alessi DR, Zhang J, Khanna A, Hochdorfer T, Shang Y, Kahle KT. The WNK-SPAK/OSR1 pathway: master regulator of cation-chloride cotransporters. Sci Signal. 2014;7:re3. https://doi.org/10.1126/scisignal.2005365
  14. Arroyo JP, Kahle KT, Gamba G. The SLC12 family of electroneutral cation-coupled chloride cotransporters. Mol Aspects Med. 2013;34:288-298. https://doi.org/10.1016/j.mam.2012.05.002
  15. Kahle KT, Rinehart J, Lifton RP. Phosphoregulation of the Na-K-2Cl and K-Cl cotransporters by the WNK kinases. Biochim Biophys Acta. 2010;1802:1150-1158. https://doi.org/10.1016/j.bbadis.2010.07.009
  16. Schumacher FR, Sorrell FJ, Alessi DR, Bullock AN, Kurz T. Structural and biochemical characterization of the KLHL3-WNK kinase interaction important in blood pressure regulation. Biochem J. 2014;460:237-246. https://doi.org/10.1042/BJ20140153
  17. McCormick JA, Yang CL, Zhang C, Davidge B, Blankenstein KI, Terker AS, Yarbrough B, Meermeier NP, Park HJ, McCully B, West M, Borschewski A, Himmerkus N, Bleich M, Bachmann S, Mutig K, Argaiz ER, Gamba G, Singer JD, Ellison DH. Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3. J Clin Invest. 2014;124:4723-4736. https://doi.org/10.1172/JCI76126
  18. Wakabayashi M, Mori T, Isobe K, Sohara E, Susa K, Araki Y, Chiga M, Kikuchi E, Nomura N, Mori Y, Matsuo H, Murata T, Nomura S, Asano T, Kawaguchi H, Nonoyama S, Rai T, Sasaki S, Uchida S. Impaired KLHL3-mediated ubiquitination of WNK4 causes human hypertension. Cell Rep. 2013;3:858-868. https://doi.org/10.1016/j.celrep.2013.02.024
  19. Takahashi D, Mori T, Wakabayashi M, Mori Y, Susa K, Zeniya M, Sohara E, Rai T, Sasaki S, Uchida S. KLHL2 interacts with and ubiquitinates WNK kinases. Biochem Biophys Res Commun. 2013;437:457-462. https://doi.org/10.1016/j.bbrc.2013.06.104
  20. Ohta A, Schumacher FR, Mehellou Y, Johnson C, Knebel A, Macartney TJ, Wood NT, Alessi DR, Kurz T. The CUL3-KLHL3 E3 ligase complex mutated in Gordon's hypertension syndrome interacts with and ubiquitylates WNK isoforms: disease-causing mutations in KLHL3 and WNK4 disrupt interaction. Biochem J. 2013;451:111-122. https://doi.org/10.1042/BJ20121903
  21. Mori Y, Wakabayashi M, Mori T, Araki Y, Sohara E, Rai T, Sasaki S, Uchida S. Decrease of WNK4 ubiquitination by disease-causing mutations of KLHL3 through different molecular mechanisms. Biochem Biophys Res Commun. 2013;439:30-34. https://doi.org/10.1016/j.bbrc.2013.08.035
  22. Hossain Khan MZ, Sohara E, Ohta A, Chiga M, Inoue Y, Isobe K, Wakabayashi M, Oi K, Rai T, Sasaki S, Uchida S. Phosphorylation of Na-Cl cotransporter by OSR1 and SPAK kinases regulates its ubiquitination. Biochem Biophys Res Commun. 2012;425:456-461. https://doi.org/10.1016/j.bbrc.2012.07.124
  23. Nishida H, Sohara E, Nomura N, Chiga M, Alessi DR, Rai T, Sasaki S, Uchida S. Phosphatidylinositol 3-kinase/Akt signaling pathway activates the WNK-OSR1/SPAK-NCC phosphorylation cascade in hyperinsulinemic db/db mice. Hypertension. 2012;60: 981-990. https://doi.org/10.1161/HYPERTENSIONAHA.112.201509
  24. Naguro I, Umeda T, Kobayashi Y, Maruyama J, Hattori K, Shimizu Y, Kataoka K, Kim-Mitsuyama S, Uchida S, Vandewalle A, Noguchi T, Nishitoh H, Matsuzawa A, Takeda K, Ichijo H. ASK3 responds to osmotic stress and regulates blood pressure by suppressing WNK1-SPAK/OSR1 signaling in the kidney. Nat Commun. 2012;3:1285. https://doi.org/10.1038/ncomms2283
  25. Kaji T, Yoshida S, Kawai K, Fuchigami Y, Watanabe W, Kubodera H, Kishimoto T. ASK3, a novel member of the apoptosis signalregulating kinase family, is essential for stress-induced cell death in HeLa cells. Biochem Biophys Res Commun. 2010;395: 213-218. https://doi.org/10.1016/j.bbrc.2010.03.164
  26. Dowd BF, Forbush B. PASK (proline-alanine-rich STE20-related kinase), a regulatory kinase of the Na-K-Cl cotransporter (NKCC1). J Biol Chem. 2003;278:27347-27353. https://doi.org/10.1074/jbc.M301899200
  27. Piala AT, Moon TM, Akella R, He H, Cobb MH, Goldsmith EJ. Chloride sensing by WNK1 involves inhibition of autophosphorylation. Sci Signal. 2014;7:ra41. https://doi.org/10.1126/scisignal.2005050
  28. Berridge MJ, Bootman MD, Roderick HL. Calcium signalling: dynamics, homeostasis and remodelling. Nat Rev Mol Cell Biol. 2003;4:517-529.
  29. Park S, Lee SI, Shin DM. Role of regulators of g-protein signaling 4 in ca signaling in mouse pancreatic acinar cells. Korean J Physiol Pharmacol. 2011;15:383-388. https://doi.org/10.4196/kjpp.2011.15.6.383
  30. Evans RL, Turner RJ. Upregulation of Na(+)-K(+)-2Cl- cotransporter activity in rat parotid acinar cells by muscarinic stimulation. J Physiol. 1997;499:351-359. https://doi.org/10.1113/jphysiol.1997.sp021932
  31. Na T, Wu G, Peng JB. Disease-causing mutations in the acidic motif of WNK4 impair the sensitivity of WNK4 kinase to calcium ions. Biochem Biophys Res Commun. 2012;419:293-298. https://doi.org/10.1016/j.bbrc.2012.02.013
  32. Hoorn EJ, Walsh SB, McCormick JA, Furstenberg A, Yang CL, Roeschel T, Paliege A, Howie AJ, Conley J, Bachmann S, Unwin RJ, Ellison DH. The calcineurin inhibitor tacrolimus activates the renal sodium chloride cotransporter to cause hypertension. Nat Med. 2011;17:1304-1309. https://doi.org/10.1038/nm.2497
  33. Diver JM, Sage SO, Rosado JA. The inositol trisphosphate receptor antagonist 2-aminoethoxydiphenylborate (2-APB) blocks $Ca^{2+}$ entry channels in human platelets: cautions for its use in studying $Ca^{2+}$ influx. Cell Calcium. 2001;30:323-329. https://doi.org/10.1054/ceca.2001.0239
  34. Bootman MD, Collins TJ, Mackenzie L, Roderick HL, Berridge MJ, Peppiatt CM. 2-aminoethoxydiphenyl borate (2-APB) is a reliable blocker of store-operated $Ca^{2+}$ entry but an inconsistent inhibitor of InsP3-induced $Ca^{2+}$ release. FASEB J. 2002;16: 1145-1150. https://doi.org/10.1096/fj.02-0037rev
  35. Hatano N, Itoh Y, Muraki K. Cardiac fibroblasts have functional TRPV4 activated by 4alpha-phorbol 12,13-didecanoate. Life Sci. 2009;85:808-814. https://doi.org/10.1016/j.lfs.2009.10.013
  36. Kim KS, Shin DH, Nam JH, Park KS, Zhang YH, Kim WK, Kim SJ. Functional expression of trpv4 cation channels in human mast cell line (HMC-1). Korean J Physiol Pharmacol. 2010;14:419-425. https://doi.org/10.4196/kjpp.2010.14.6.419
  37. Nilius B, Vriens J, Prenen J, Droogmans G, Voets T. TRPV4 calcium entry channel: a paradigm for gating diversity. Am J Physiol Cell Physiol. 2004;286:C195-205. https://doi.org/10.1152/ajpcell.00365.2003
  38. Kline D, Kline JT. Repetitive calcium transients and the role of calcium in exocytosis and cell cycle activation in the mouse egg. Dev Biol. 1992;149:80-89. https://doi.org/10.1016/0012-1606(92)90265-I
  39. Melvin JE, Yule D, Shuttleworth T, Begenisich T. Regulation of fluid and electrolyte secretion in salivary gland acinar cells. Annu Rev Physiol. 2005;67:445-469. https://doi.org/10.1146/annurev.physiol.67.041703.084745
  40. Wu Y, Schellinger JN, Huang CL, Rodan AR. Hypotonicity stimulates potassium flux through the WNK-SPAK/OSR1 kinase cascade and the Ncc69 sodium-potassium-2-chloride cotransporter in the Drosophila renal tubule. J Biol Chem. 2014;289:26131-26142. https://doi.org/10.1074/jbc.M114.577767

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