Abstract
Organic anion transporter 3 (OAT3) plays a vital role in removing a broad variety of anionic drugs from kidney, thus avoiding their possible toxicity in the body. We earlier established that activation of protein kinase C (PKC) enhances OAT3 ubiquitination, which promotes OAT3 internalization from the cell plasma membrane to intracellular endosomes and consequent degradation. As a result, OAT3 expression and transport activity are reduced. In the current study, we discovered that protein kinase A (PKA) had an opposite effect to PKC on the regulation of OAT3. We showed that activation of PKA by Bt2-cAMP stimulated OAT3 transport activity, which was largely caused by an enhanced plasma membrane expression of the transporter, kinetically reflected as an augmented maximal transport velocity Vmax without notable alteration in substrate-binding affinity Km. Additionally, we showed that PKA activation accelerated the rate of OAT3 recycling from intracellular compartments to the plasma membrane and decelerated the rate of OAT3 degradation. We further showed that OAT3 is subjected to post-translational modification by SUMO-2 and SUMO-3 not by SUMO-1. PKA activation enhanced OAT3 SUMOylation, which was accompanied by a reduced OAT3 ubiquitination. Finally, insulin-like growth factor 1 significantly stimulated OAT3 transport activity and SUMOylation through PKA signaling pathway. In conclusion, this is the first demonstration that PKA stimulated OAT3 expression and transport activity by altering the trafficking kinetics of OAT3 possibly through the crosstalk between SUMOylation and ubiquitination. Our studies are consistent with a remote sensing and signaling model for transporters (Wu et al. in Mol Pharmacol. 79(5):795–805, 2011).
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References
You G. Structure, function, and regulation of renal organic anion transporters. Med Res Rev. 2002;22(6):602–16. https://doi.org/10.1002/med.10019.
Srimaroeng C, Perry JL, Pritchard JB. Physiology, structure, and regulation of the cloned organic anion transporters. Xenobiotica. 2008;38(7–8):889–935. https://doi.org/10.1080/00498250801927435.
Dantzler WH, Wright SH. The molecular and cellular physiology of basolateral organic anion transport in mammalian renal tubules. Biochim Biophys Acta. 2003;1618(2):185–93.
VanWert AL, Gionfriddo MR, Sweet DH. Organic anion transporters: discovery, pharmacology, regulation and roles in pathophysiology. Biopharm Drug Dispos. 2010;31(1):1–71. https://doi.org/10.1002/bdd.693.
Ahn SY, Nigam SK. Toward a systems level understanding of organic anion and other multispecific drug transporters: a remote sensing and signaling hypothesis. Mol Pharmacol. 2009;76(3):481–90. https://doi.org/10.1124/mol.109.056564.
Terada T, Inui K. Gene expression and regulation of drug transporters in the intestine and kidney. Biochem Pharmacol. 2007;73(3):440–9. https://doi.org/10.1016/j.bcp.2006.10.010.
Zhang Q, Hong M, Duan P, Pan Z, Ma J, You G. Organic anion transporter OAT1 undergoes constitutive and protein kinase C-regulated trafficking through a dynamin- and clathrin-dependent pathway. J Biol Chem. 2008;283(47):32570–9. https://doi.org/10.1074/jbc.M800298200.
Zhang Q, Pan Z, You G. Regulation of human organic anion transporter 4 by protein kinase C and NHERF-1: altering the endocytosis of the transporter. Pharm Res. 2010;27(4):589–96. https://doi.org/10.1007/s11095-009-9983-2.
Zhang Q, Suh W, Pan Z, You G. Short-term and long-term effects of protein kinase C on the trafficking and stability of human organic anion transporter 3. Int J Biochem Mol Biol. 2012;3(2):242–9.
Xu D, Zhang J, Zhang Q, Fan Y, Liu C, You G. PKC/Nedd4-2 signaling pathway regulates the cell surface expression of drug transporter hOAT1. Drug Metab Dispos. 2017;45(8):887–95. https://doi.org/10.1124/dmd.117.075861.
Gareau JR, Lima CD. The SUMO pathway: emerging mechanisms that shape specificity, conjugation and recognition. Nat Rev Mol Cell Biol. 2010;11(12):861–71. https://doi.org/10.1038/nrm3011.
Ulrich HD. The fast-growing business of SUMO chains. Mol Cell. 2008;32(3):301–5. https://doi.org/10.1016/j.molcel.2008.10.010.
Plant LD, Dementieva IS, Kollewe A, Olikara S, Marks JD, Goldstein SA. One SUMO is sufficient to silence the dimeric potassium channel K2P1. Proc Natl Acad Sci U S A. 2010;107(23):10743–8. https://doi.org/10.1073/pnas.1004712107.
Rajan S, Plant LD, Rabin ML, Butler MH, Goldstein SA. Sumoylation silences the plasma membrane leak K+ channel K2P1. Cell. 2005;121(1):37–47. https://doi.org/10.1016/j.cell.2005.01.019.
Rajan S, Dickson LM, Mathew E, Orr CM, Ellenbroek JH, Philipson LH, et al. Chronic hyperglycemia downregulates GLP-1 receptor signaling in pancreatic beta-cells via protein kinase A. Mol Metab. 2015;4(4):265–76. https://doi.org/10.1016/j.molmet.2015.01.010.
Kang JS, Saunier EF, Akhurst RJ, Derynck R. The type I TGF-beta receptor is covalently modified and regulated by sumoylation. Nat Cell Biol. 2008;10(6):654–64. https://doi.org/10.1038/ncb1728.
Tang Z, El Far O, Betz H, Scheschonka A. Pias1 interaction and sumoylation of metabotropic glutamate receptor 8. J Biol Chem. 2005;280(46):38153–9. https://doi.org/10.1074/jbc.M508168200.
Minami S, Ito K, Honma M, Ikebuchi Y, Anzai N, Kanai Y, et al. Posttranslational regulation of Abcc2 expression by SUMOylation system. Am J Physiol Gastrointest Liver Physiol. 2009;296(2):G406–13. https://doi.org/10.1152/ajpgi.90309.2008.
Giorgino F, de Robertis O, Laviola L, Montrone C, Perrini S, McCowen KC, et al. The sentrin-conjugating enzyme mUbc9 interacts with GLUT4 and GLUT1 glucose transporters and regulates transporter levels in skeletal muscle cells. Proc Natl Acad Sci U S A. 2000;97(3):1125–30.
Ulrich HD. Mutual interactions between the SUMO and ubiquitin systems: a plea of no contest. Trends Cell Biol. 2005;15(10):525–32. https://doi.org/10.1016/j.tcb.2005.08.002.
Desterro JM, Rodriguez MS, Hay RT. SUMO-1 modification of IkappaBalpha inhibits NF-kappaB activation. Mol Cell. 1998;2(2):233–9.
Duan P, Li S, You G. Regulation of human organic anion transporter 4 by parathyroid hormone-related protein and protein kinase A. Int J Biochem Mol Biol. 2012;3(3):322–7.
Barros SA, Srimaroeng C, Perry JL, Walden R, Dembla-Rajpal N, Sweet DH, et al. Activation of protein kinase Czeta increases OAT1 (SLC22A6)- and OAT3 (SLC22A8)-mediated transport. J Biol Chem. 2009;284(5):2672–9. https://doi.org/10.1074/jbc.M808078200.
Soodvilai S, Chatsudthipong V, Evans KK, Wright SH, Dantzler WH. Acute regulation of OAT3-mediated estrone sulfate transport in isolated rabbit renal proximal tubules. Am J Physiol Ren Physiol. 2004;287(5):F1021–9. https://doi.org/10.1152/ajprenal.00080.2004.
Subramaniam S, Shahani N, Strelau J, Laliberte C, Brandt R, Kaplan D, et al. Insulin-like growth factor 1 inhibits extracellular signal-regulated kinase to promote neuronal survival via the phosphatidylinositol 3-kinase/protein kinase A/c-Raf pathway. J Neurosci. 2005;25(11):2838–52. https://doi.org/10.1523/JNEUROSCI.5060-04.2005.
Wang H, You G. SGK1/Nedd4-2 signaling pathway regulates the activity of human organic anion transporters 3. Biopharm Drug Dispos. 2017;38(8):449–57. https://doi.org/10.1002/bdd.2085.
Wang H, Xu D, Toh MF, Pao AC, You G. Serum- and glucocorticoid-inducible kinase SGK2 regulates human organic anion transporters 4 via ubiquitin ligase Nedd4-2. Biochem Pharmacol. 2016;102:120–9. https://doi.org/10.1016/j.bcp.2015.11.024.
Zhang Q, Li S, Patterson C, You G. Lysine 48-linked polyubiquitination of organic anion transporter-1 is essential for its protein kinase C-regulated endocytosis. Mol Pharmacol. 2013;83(1):217–24. https://doi.org/10.1124/mol.112.082065.
Duan P, Li S, You G. Angiotensin II inhibits activity of human organic anion transporter 3 through activation of protein kinase Calpha: accelerating endocytosis of the transporter. Eur J Pharmacol. 2010;627(1–3):49–55. https://doi.org/10.1016/j.ejphar.2009.10.048.
Phatchawan A, Chutima S, Varanuj C, Anusorn L. Decreased renal organic anion transporter 3 expression in type 1 diabetic rats. Am J Med Sci. 2014;347(3):221–7. https://doi.org/10.1097/MAJ.0b013e3182831740.
Muller S, Matunis MJ, Dejean A. Conjugation with the ubiquitin-related modifier SUMO-1 regulates the partitioning of PML within the nucleus. EMBO J. 1998;17(1):61–70. https://doi.org/10.1093/emboj/17.1.61.
Yang SH, Jaffray E, Senthinathan B, Hay RT, Sharrocks AD. SUMO and transcriptional repression: dynamic interactions between the MAP kinase and SUMO pathways. Cell Cycle. 2003;2(6):528–30. https://doi.org/10.4161/cc.2.6.597.
Hietakangas V, Ahlskog JK, Jakobsson AM, Hellesuo M, Sahlberg NM, Holmberg CI, et al. Phosphorylation of serine 303 is a prerequisite for the stress-inducible SUMO modification of heat shock factor 1. Mol Cell Biol. 2003;23(8):2953–68.
Bach LA, Hale LJ. Insulin-like growth factors and kidney disease. Am J Kidney Dis. 2015;65(2):327–36. https://doi.org/10.1053/j.ajkd.2014.05.024.
Yakar S, Adamo ML. Insulin-like growth factor 1 physiology: lessons from mouse models. Endocrinol Metab Clin N Am. 2012;41(2):231–47, v. https://doi.org/10.1016/j.ecl.2012.04.008.
Giustina A, Chanson P, Kleinberg D, Bronstein MD, Clemmons DR, Klibanski A, et al. Expert consensus document: a consensus on the medical treatment of acromegaly. Nat Rev Endocrinol. 2014;10(4):243–8. https://doi.org/10.1038/nrendo.2014.21.
Janecka A, Kolodziej-Rzepa M, Biesaga B. Clinical and molecular features of Laron syndrome, a genetic disorder protecting from cancer. In Vivo. 2016;30(4):375–81.
Bang P, Polak M, Woelfle J, Houchard A, Group EIRS. Effectiveness. Safety of rhIGF-1 therapy in children: The European Increlex(R) Growth Forum Database Experience. Horm Res Paediatr. 2015;83(5):345–57. https://doi.org/10.1159/000371798.
Cohen J, Blethen S, Kuntze J, Smith SL, Lomax KG, Mathew PM. Managing the child with severe primary insulinlike growth factor-1 deficiency (IGFD): IGFD diagnosis and management. Drugs R D. 2014;14(1):25–9. https://doi.org/10.1007/s40268-014-0039-7.
Wu W, Dnyanmote AV, Nigam SK. Remote communication through solute carriers and ATP binding cassette drug transporter pathways: an update on the remote sensing and signaling hypothesis. Mol Pharmacol. 2011;79(5):795–805. https://doi.org/10.1124/mol.110.070607.
Bush KT, Wu W, Lun C, Nigam SK. The drug transporter OAT3 (SLC22A8) and endogenous metabolite communication via the gut-liver-kidney axis. J Biol Chem. 2017;292(38):15789–803. https://doi.org/10.1074/jbc.M117.796516.
Wu W, Jamshidi N, Eraly SA, Liu HC, Bush KT, Palsson BO, et al. Multispecific drug transporter Slc22a8 (Oat3) regulates multiple metabolic and signaling pathways. Drug Metab Dispos. 2013;41(10):1825–34. https://doi.org/10.1124/dmd.113.052647.
Acknowledgments
We would like to thank Dr. Jorge A Iñiguez-Lluhí for his generous gifts of HA-tagged SUMO1, SUMO2, SUMO3, and Ubc9 plasmids.
Funding
This work was supported by grants (to Dr. Guofeng You) from National Institute of General Medical Sciences (R01-GM079123 and R01-GM097000).
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Wang, H., Zhang, J. & You, G. Activation of Protein Kinase A Stimulates SUMOylation, Expression, and Transport Activity of Organic Anion Transporter 3. AAPS J 21, 30 (2019). https://doi.org/10.1208/s12248-019-0303-4
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DOI: https://doi.org/10.1208/s12248-019-0303-4