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Scaffolding protein GOPC regulates tight junction structure

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Abstract

GOPC (FIG/PIST/CAL) is a PDZ-domain scaffolding protein that regulates the trafficking of a wide array of proteins, including small GTPases, receptors and cell surface molecules such as cadherin 23 and cystic fibrosis transmembrane regulator. In Madin-Darby canine kidney (MDCK) cells, we find that GOPC localizes to the trans-Golgi network (TGN) but not to the cis- or trans-Golgi cisternae. Colocalization occurs with the early endosome Rab GTPase Rab5 and a TGN/endosome marker Rab14 but not with Rab11, a marker of recycling endosomes. No localization of GOPC was detected to the lateral membranes or tight junctions. Knockdown of GOPC in MDCK cells results in decreased transepithelial resistance and increased paracellular flux. This might be attributable to the compromised trafficking of tight junction components from the TGN, as GOPC-knockdown cells have decreased lateral labeling of the tight junction protein claudin-1 and decreased protein levels of claudin-2. GOPC might mediate the trafficking of newly synthesized tight junction proteins from the TGN to the cell surface or the recycling of these proteins from specialized endosomal compartments.

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Acknowledgments

We thank members of the Wilson laboratory for helpful discussions and the Delamere and Simon laboratories for assistance with the dextran flux assays. We also thank Dr. Charles Parkos for antibody against JAM-A and the Broad Institute for small hairpin RNA against GOPC.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, University of Arizona, PO Box 245044, Tucson, AZ 85724, USA

    Ruifeng Lu, Lorraine Stewart & Jean M. Wilson

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  1. Ruifeng Lu
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  2. Lorraine Stewart
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  3. Jean M. Wilson
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Corresponding author

Correspondence to Jean M. Wilson.

Additional information

This work was supported by NIH RO1 DK84047 (to J.M.W.).

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Supplemental Figure 1

Normal rat kidney (NRK) cells labeled for TGN38 (a) and GOPC (a’). The merged image shows that endogenous TGN38 co-localizes with GOPC (a’’). (GIF 39 kb)

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Lu, R., Stewart, L. & Wilson, J.M. Scaffolding protein GOPC regulates tight junction structure. Cell Tissue Res 360, 321–332 (2015). https://doi.org/10.1007/s00441-014-2088-1

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