Human Molecular Genetics, 2002, Vol. 11, No. 1 59-67
© 2002 Oxford University Press
Distinct subcellular expression of endogenous polycystin-2 in the plasma membrane and Golgi apparatus of MDCK cells
Department of Human and Clinical Genetics, Sylvius Laboratory, Leiden University Medical Center, 2333AL Leiden, The Netherlands and 1Department of Pathology, Leiden University Medical Center, 2333ZA Leiden, The Netherlands
Polycystin-2 is a predicted integral membrane protein with non-selective cation channel activity. The protein is encoded by the PKD2 gene, which is mutated in ~15% of patients with autosomal dominant polycystic kidney disease (ADPKD). Polycystin-2 can interact with the transmembrane protein polycystin-1, the product of the PKD1 gene. However, endoplasmic reticulum (ER) localization was reported for (heterologously expressed) polycystin-2 in cultured cells and baso-lateral localization has been reported in renal tissues. Using two polyclonal antisera raised against polycystin-2 we demonstrated distinct expression of the endogenous protein in the Golgi apparatus and the plasma membrane of MDCK cells. In contrast, most of the heterologously expressed polycystin-2 (PC2–EGFP) remained in the ER, substantially overlapping with the staining pattern of protein-disulfide isomerase (PDI), a marker for the ER. Only in a small subset of these cells weak plasma membrane signals were observed. Membrane staining was also suggested by immunoelectron microscopy and was confirmed by subcellular fractionation on sucrose density gradients. The plasma membrane staining disappeared following extraction with a buffer containing Triton X-100, whereas signals for polycystin-1 and E-cadherin remained visible, suggesting that polycystin-2 is neither tightly bound to the Triton X-100 insoluble cytoskeleton, nor to these proteins. We conclude that endogenous polycystin-2 is transported via the Golgi apparatus to the plasma membrane and has a broader membrane localization than polycystin-1. These data suggest that polycystin-2 can move freely in certain regions of the membrane where it probably functions as a channel, activated by, or in complex with, polycystin-1.
+ To whom correspondence should be addressed. Tel: +31 71 5276048; Fax: +31 71 5276075; Email: d.j.m.peters@lumc.nl
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  V. M. Ripoll, K. M. Irvine, T. Ravasi, M. J. Sweet, and D. A. Hume Gpnmb Is Induced in Macrophages by IFN-{gamma} and Lipopolysaccharide and Acts as a Feedback Regulator of Proinflammatory Responses J. Immunol., May 15, 2007; 178(10): 6557 - 6566. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M. Turner, B. F. King, K. S. Srai, and R. J. Unwin Antagonism of endogenous putative P2Y receptors reduces the growth of MDCK-derived cysts cultured in vitro Am J Physiol Renal Physiol, January 1, 2007; 292(1): F15 - F25. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Obara, S. Mangos, Y. Liu, J. Zhao, S. Wiessner, A. G. Kramer-Zucker, F. Olale, A. F. Schier, and I. A. Drummond Polycystin-2 Immunolocalization and Function in Zebrafish J. Am. Soc. Nephrol., October 1, 2006; 17(10): 2706 - 2718. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E.-F. Bui-Xuan, Q. Li, X.-Z. Chen, C. A. Boucher, R. Sandford, J. Zhou, and N. Basora More than colocalizing with polycystin-1, polycystin-L is in the centrosome Am J Physiol Renal Physiol, August 1, 2006; 291(2): F395 - F406. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Shiratori and H. Hamada The left-right axis in the mouse: from origin to morphology Development, June 1, 2006; 133(11): 2095 - 2104. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Streets, D. J. Moon, M. E. Kane, T. Obara, and A. C.M. Ong Identification of an N-terminal glycogen synthase kinase 3 phosphorylation site which regulates the functional localization of polycystin-2 in vivo and in vitro Hum. Mol. Genet., May 1, 2006; 15(9): 1465 - 1473. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Li, N. Montalbetti, P. Y. Shen, X.-Q. Dai, C. I. Cheeseman, E. Karpinski, G. Wu, H. F. Cantiello, and X.-Z. Chen Alpha-actinin associates with polycystin-2 and regulates its channel activity Hum. Mol. Genet., June 15, 2005; 14(12): 1587 - 1603. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Pluznick, P. Wei, P. R. Grimm, and S. C. Sansom BK-{beta}1 subunit: immunolocalization in the mammalian connecting tubule and its role in the kaliuretic response to volume expansion Am J Physiol Renal Physiol, April 1, 2005; 288(4): F846 - F854. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Jurczyk, A. Gromley, S. Redick, J. S. Agustin, G. Witman, G. J. Pazour, D. J.M. Peters, and S. Doxsey Pericentrin forms a complex with intraflagellar transport proteins and polycystin-2 and is required for primary cilia assembly J. Cell Biol., August 30, 2004; 166(5): 637 - 643. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Zhang, P. D. Taulman, and B. K. Yoder Cystic Kidney Diseases: All Roads Lead to the Cilium Physiology, August 1, 2004; 19(4): 225 - 230. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. R. Rundle, G. Gorbsky, and L. Tsiokas PKD2 Interacts and Co-localizes with mDia1 to Mitotic Spindles of Dividing Cells: ROLE OF mDia1 IN PKD2 LOCALIZATION TO MITOTIC SPINDLES J. Biol. Chem., July 9, 2004; 279(28): 29728 - 29739. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. H. Le, P. van der Bent, G. Huls, M. van de Wetering, M. Loghman-Adham, A. C. M. Ong, J. P. Calvet, H. Clevers, M. H. Breuning, H. van Dam, et al. Aberrant Polycystin-1 Expression Results in Modification of Activator Protein-1 Activity, whereas Wnt Signaling Remains Unaffected J. Biol. Chem., June 25, 2004; 279(26): 27472 - 27481. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Babich, W.-Z. Zeng, B.-I. Yeh, O. Ibraghimov-Beskrovnaya, Y. Cai, S. Somlo, and C.-L. Huang The N-terminal Extracellular Domain Is Required for Polycystin-1-dependent Channel Activity J. Biol. Chem., June 11, 2004; 279(24): 25582 - 25589. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Gao, E. Joseph, D. M. Ruden, and X. Lu Drosophila Pkd2 Is Haploid-insufficient for Mediating Optimal Smooth Muscle Contractility J. Biol. Chem., April 2, 2004; 279(14): 14225 - 14231. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. W. M. van Balkom, M. P. J. Graat, M. van Raak, E. Hofman, P. van der Sluijs, and P. M. T. Deen Role of cytoplasmic termini in sorting and shuttling of the aquaporin-2 water channel Am J Physiol Cell Physiol, February 1, 2004; 286(2): C372 - C379. [Abstract] [Full Text]
|
|
|
|
 |
|
 D. Joly, A. Hummel, A. Ruello, and B. Knebelmann Ciliary function of polycystins: a new model for cystogenesis Nephrol. Dial. Transplant., September 1, 2003; 18(9): 1689 - 1692. [Full Text] [PDF]
|
|
|
|
|
|
Q. Qian, M. Li, Y. Cai, C. J. Ward, S. Somlo, P. C. Harris, and V. E. Torres Analysis of the Polycystins in Aortic Vascular Smooth Muscle Cells J. Am. Soc. Nephrol., September 1, 2003; 14(9): 2280 - 2287. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Qian, L. W. Hunter, M. Li, M. Marin-Padilla, Y.S. Prakash, S. Somlo, P. C. Harris, V. E. Torres, and G. C. Sieck Pkd2 haploinsufficiency alters intracellular calcium regulation in vascular smooth muscle cells Hum. Mol. Genet., August 1, 2003; 12(15): 1875 - 1880. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Calvet New insights into ciliary function: Kidney cysts and photoreceptors PNAS, May 13, 2003; 100(10): 5583 - 5585. [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Luo, P. M. Vassilev, X. Li, Y. Kawanabe, and J. Zhou Native Polycystin 2 Functions as a Plasma Membrane Ca2+-Permeable Cation Channel in Renal Epithelia Mol. Cell. Biol., April 1, 2003; 23(7): 2600 - 2607. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Zhao, J. L. Haylor, and A. C. M. Ong Polycystin-2 expression is increased following experimental ischaemic renal injury Nephrol. Dial. Transplant., December 1, 2002; 17(12): 2138 - 2144. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. K. Yoder, X. Hou, and L. M. Guay-Woodford The Polycystic Kidney Disease Proteins, Polycystin-1, Polycystin-2, Polaris, and Cystin, Are Co-Localized in Renal Cilia J. Am. Soc. Nephrol., October 1, 2002; 13(10): 2508 - 2516. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Calvet Cilia in PKD--Letting It All Hang Out J. Am. Soc. Nephrol., October 1, 2002; 13(10): 2614 - 2616. [Full Text] [PDF]
|
|
|
|
|
|
P. Igarashi and S. Somlo Genetics and Pathogenesis of Polycystic Kidney Disease J. Am. Soc. Nephrol., September 1, 2002; 13(9): 2384 - 2398. [Full Text] [PDF]
|
|