Inhibition of endocytosis causes phosphorylation (S256)-independent plasma membrane accumulation of AQP2

American Journal of Physiology - Renal Physiology - Tập 286 Số 2 - Trang F233-F243 - 2004
Hua Lu1, Tian‐Xiao Sun1, Richard Bouley1, Karen Blackburn1, Margaret McLaughlin1, Dennis Brown1
1Program in Membrane Biology and Renal Unit, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114

Tóm tắt

Inhibition of clathrin-mediated endocytosis by expression of a GTPase-deficient dynamin mutant (dynamin-2/K44A) for 16 h results in an accumulation of plasma membrane aquaporin-2 (AQP2) in epithelial cells stably transfected with wild-type AQP2. We now show a similar effect of K44A dynamin in LLC-PK1cells transfected with an S256 phosphorylation-deficient AQP2 mutant, AQP2(S256A), and in AQP2-transfected inner medullary collecting duct (IMCD) cells. More acute blockade of endocytosis in these cells with the cholesterol-depleting agent methyl-β-cyclodextrin (mβCD; 10 mM) resulted in a rapid and extensive cell-surface accumulation of both wild-type AQP2 and AQP2 (S256A) within 15 min after treatment. This effect was similar to that induced by treatment of the cells with vasopressin. Blockade of endocytosis by mβCD was confirmed using quantitative analysis of FITC-dextran uptake and AQP2 membrane insertion was verified by cell-surface biotinylation. These data indicate that AQP2 recycles constitutively and rapidly between intracellular stores and the cell surface in LLC-PK1 and IMCD cells. The constitutive trafficking process is not dependent on phosphorylation of the serine-256 residue of AQP2, which is, however, an essential step for regulated vasopressin/cAMP-mediated translocation of AQP2. Our data show that rapid and extensive plasma membrane accumulation of AQP2 can occur in a vasopressin receptor (V2R)- and phosphorylation-independent manner, pointing to a potential means of bypassing the mutated V2R in X-linked nephrogenic diabetes insipidus to achieve cell surface expression of AQP2.

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Tài liệu tham khảo

10.1074/jbc.273.28.17504

10.1172/JCI9594

10.1152/ajprenal.00387.2002

Brown Dand Nielsen S.Cell biology of vasopressin action. In:The Kidney, edited by Brenner BM. Philadelphia, PA: Saunders, 2000, p. 575-594.

10.1038/302253a0

Brown D, Weyer P, and Orci L.Vasopressin stimulates endocytosis in kidney collecting duct principal cells.Eur J Cell Biol46: 336-341, 1988.

10.1073/pnas.091502398

10.1074/jbc.M005552200

10.1152/ajprenal.2000.278.1.F29

Deen PM, Rijss JP, Mulders SM, Errington RJ, van Baal J, and van Os CH.Aquaporin-2 transfection of Madin-Darby canine kidney cells reconstitutes vasopressin-regulated transcellular osmotic water transport.J Am Soc Nephrol8: 1493-1501, 1997.

Deen PMTand Brown D.Trafficking of native and mutant mammalian MIP proteins. In:Aquaporins: Current Topics in Membranes, edited by Hohmann S, Nielsen S, and Agre P. New York: Academic, 2001, p. 235-276.

10.1074/jbc.272.23.14800

10.1152/ajprenal.2000.278.2.F317

10.1007/BF00852577

10.1146/annurev.cellbio.16.1.483

10.1073/pnas.241368698

10.1083/jcb.151.4.919

10.1006/bbrc.1993.2111

10.1074/jbc.273.39.25450

Katsura T, Gustafson CE, Ausiello DA, and Brown D.Protein kinase A phosphorylation is involved in regulated exocytosis of aquaporin-2 in transfected LLC-PK1 cells.Am J Physiol Renal Physiol272: F817-F822, 1997.

10.1073/pnas.92.16.7212

10.1074/jbc.M010270200

10.1021/bi0007021

10.1016/S0968-0004(99)01538-8

10.1146/annurev.physiol.63.1.607

10.1152/physrev.00024.2001

10.1152/ajprenal.1999.276.2.F254

10.1042/bj3460321

10.1006/bbrc.1997.7810

10.1091/mbc.10.4.961

10.1093/ndt/15.suppl_6.21

Satoh S, Nishimura H, Clark AE, Kozka IJ, Vannucci SJ, Simpson IA, Quon MJ, Cushman SW, and Holman GD.Use of bismannose photo-label to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action.J Biol Chem268: 17820-17829, 1993.

10.1034/j.1600-0854.2000.010503.x

10.1083/jcb.145.4.877

10.1091/mbc.E02-06-0315

10.1074/jbc.M208563200

10.1007/BF01871929

10.1073/pnas.96.12.6775

10.1152/ajprenal.00257.2001

10.1016/S0960-9822(02)01223-X

Thorens Band Roth J.Intracellular targeting of GLUT4 in transfected insulinoma cells: evidence for association with constitutively recycling vesicles distinct from synaptophysin and insulin vesicles.J Cell Sci109: 1311-1323, 1996.

Valenti G, Procino G, Carmosino M, Frigeri A, Mannucci R, Nicoletti I, and Svelto M.The phosphatase inhibitor okadaic acid induces AQP2 translocation independently from AQP2 phosphorylation in renal collecting duct cells.J Cell Sci113: 1985-1992, 2000.

10.1074/jbc.M207525200

10.1038/333268a0

Yang Jand Holman GD.Comparison of GLUT4 and GLUT1 subcellular trafficking in basal and insulin-stimulated 3T3-L1 cells.J Biol Chem268: 4600-4603, 1993.

10.1152/ajprenal.2000.278.3.F388

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American Journal of Physiology - Renal Physiology

Tập 286 Số 2

F233-F243

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