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Human Molecular Genetics, 2000, Vol. 9, No. 18 2743-2750
© 2000 Oxford University Press

Polycystin-1, the product of the polycystic kidney disease 1 gene, co-localizes with desmosomes in MDCK cells

Martijn S. Scheffers, Paola van der Bent, Frans Prins1, Lia Spruit, Martijn H. Breuning, Sergey V. Litvinov1, Emile de Heer1 and Dorien J.M. Peters+

Department of Human and Clinical Genetics and 1Department of Pathology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands

Polycystin-1 is a novel protein predicted to be a large membrane-spanning glycoprotein with an extracellular N-terminus and an intracellular C-terminus, harboring several structural motifs. To study the subcellular localization, antibodies raised against various domains of polycystin-1 and against specific adhesion complex proteins were used for two-color immunofluorescence staining. In Madine Darby canine kidney (MDCK) cells, polycystin-1 was detected in the cytoplasm as well as co-localizing with desmosomes, but not with tight or adherens junctions. Using confocal laser scanning and immunoelectron microscopy we confirmed the desmosomal localization. By performing a calcium switch experiment, we demonstrated the sequential reassembly of tight junctions, subsequently adherens junctions and finally desmosomes. Polycystin-1 only stained the membrane after incorporation of desmoplakin into the desmosomes, suggesting that membrane-bound polycystin-1 may be important for cellular signaling or cell adhesion, but not for the assembly of adhesion complexes.

+ To whom correspondence should be addressed. Tel: +31 715 276 048; Fax: +31 75 276 075; Email: d.j.m.peters@lumc.nl


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