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Densin and beta-catenin form a complex and co-localize in cultured podocyte cell junctions

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Abstract

Densin is a member of LAP (leucine-rich repeat and PDZ domain) protein family that localizes in kidney to slit diaphragms, which are essential components of the glomerular filtration barrier. We have previously shown that densin interacts with a crucial slit diaphragm protein, nephrin. Here, we searched for novel binding partners of densin by yeast-two hybrid assay and identified beta-catenin. The interaction was confirmed by reciprocal co-immunoprecipitation assay and the binding site in densin was determined by GST-pull down assays. The GST-tagged densin was also able to pull down P-cadherin together with beta-catenin from human kidney glomerular lysates. Furthermore, densin co-localized with beta-catenin and F-actin in cell–cell contacts in cultured mouse podocytes. During cell–cell contact disruption and reformation densin and beta-catenin were dislocated from and relocated back to plasma membrane in a similar fashion. These and our previous findings suggest that densin may associate with the cadherin-catenin and nephrin complex(es), and may be involved in the formation of the cell–cell contacts including the slit diaphragm.

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Acknowledgements

We are thankful to Claudia Kocksch for the skilled technical assistance and to Lauri Ylimaa for graphical work. Drs Masaki Inagaki and Ichiro Izawa are acknowledged for providing us with densin antibody and full-length human densin construct. This study was supported by the European Union (grants LSHB-CT-2003–503364, QLRT–2001 – 01215), Paavo Nurmi Foundation, the Academy of Finland (#213692), Finnish Diabetes Research foundation, The Sigrid Juselius Foundation and Finnish Heart Association.

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Authors and Affiliations

  1. Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland

    Eija Heikkilä, Mervi Ristola, Sanna Lehtonen, Markus Lassila, Marika Havana & Harry Holthöfer

  2. Department of Anatomy, University of Greifswald, Greifswald, Germany

    Karlhans Endlich & Nicole Endlich

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  1. Eija Heikkilä
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Correspondence to Harry Holthöfer.

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Heikkilä, E., Ristola, M., Endlich, K. et al. Densin and beta-catenin form a complex and co-localize in cultured podocyte cell junctions. Mol Cell Biochem 305, 9–18 (2007). https://doi.org/10.1007/s11010-007-9522-6

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