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Lehtonen, S, Ora, A, Olkkonen, VM, Geng, L, Zerial, M, Somlo, S and Lehtonen, E (2000) In vivo interaction of the adapter protein CD2-associated protein with the type 2 polycystic kidney disease protein, polycystin-2. J. Biol. Chem. 275:32888-93
We identified a developmentally regulated gene from mouse kidney whose expression is up-regulated in metanephrogenic mesenchyme cells when they are induced to differentiate to epithelial cells during kidney organogenesis. The deduced 70.5-kDa protein, originally named METS-1 (mesenchyme-to-epithelium transition protein with SH3 domains), has since been cloned as a CD2-associated protein (CD2AP). CD2AP is strongly expressed in glomerular podocytes, and the absence of CD2AP in mice results in congenital nephrotic syndrome. We have found that METS-1/CD2AP (hereafter referred to as CD2AP) is expressed at lower levels in renal tubular epithelial cells in the adult kidney, particularly in distal nephron segments. Independent yeast two-hybrid screens using the COOH-terminal region of either CD2AP or polycystin-2 as bait identified the COOH termini of polycystin-2 and CD2AP, respectively, as strong interacting partners. This interaction was confirmed in cultured cells by co-immunoprecipitation of endogenous polycystin-2 with endogenous CD2AP and vice versa. CD2AP shows a diffuse reticular cytoplasmic and perinuclear pattern of distribution, similar to polycystin-2, in cultured cells, and the two proteins co-localize by indirect double immunofluorescence microscopy. CD2AP is an adapter molecule that associates with a variety of membrane proteins to organize the cytoskeleton around a polarized site. Such a function fits well with that hypothesized for the polycystin proteins in renal tubular epithelial cells, and the present findings suggest that CD2AP has a role in polycystin-2 function.
Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Calcium-Binding Proteins/genetics; Calcium-Binding Proteins/metabolism; Cell Line; Cloning, Molecular; Cytoskeletal Proteins; Gene Expression Regulation, Developmental; Kidney/embryology; Kidney/growth & development; Kidney/metabolism; Kidney Tubules/metabolism; Membrane Proteins/chemistry; Membrane Proteins/genetics; Membrane Proteins/metabolism; Mice; Molecular Sequence Data; Nephrons/metabolism; Open Reading Frames; Polycystic Kidney Diseases/genetics; Proteins/chemistry; Proteins/genetics; Proteins/metabolism; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Saccharomyces cerevisiae; TRPP Cation Channels; Transfection; Urothelium/metabolism; src Homology Domains
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