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Tricellulin forms homomeric and heteromeric tight junctional complexes

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Abstract

Sealing of the paracellular cleft by tight junctions is of central importance for epithelia and endothelia to function as efficient barriers between the extracellular space and the inner milieu. Occludin and claudins represent the major tight junction components involved in establishing this barrier function. A special situation emerges at sites where three cells join together. Tricellulin, a recently identified tetraspan protein concentrated at tricellular contacts, was reported to organize tricellular as well as bicellular tight junctions. Here we show that in MDCK cells, the tricellulin C-terminus is important for the basolateral translocation of tricellulin, whereas the N-terminal domain appears to be involved in directing tricellulin to tricellular contacts. In this respect, identification of homomeric tricellulin-tricellulin and of heteromeric tricellulin-occludin complexes extends a previously published model and suggests that tricellulin and occludin are transported together to the edges of elongating bicellular junctions and get separated when tricellular contacts are formed.

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Acknowledgments

This work was supported by the DFG Research Group FOR 721 and the Sonnenfeld-Stiftung. We thank Dr. Michael Schaefer for providing pcDNA3-NYFP, pcDNA3-NCFP, pcDNA3-CYFP and pcDNA3-CCFP vectors and Luise Kosel for technical assistance.

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Authors and Affiliations

  1. Institute of Biochemistry II, Jena University Hospital, Nonnenplan 2, 07743, Jena, Germany

    Julie K. Westphal, Max J. Dörfel & Otmar Huber

  2. Institute of Laboratory Medicine and Pathobiochemistry, Charité-Universitätsmedizin Berlin, Berlin, Germany

    Julie K. Westphal, Max J. Dörfel, Rudolf Tauber & Otmar Huber

  3. Institute of Clinical Physiology, Charité-Universitätsmedizin Berlin, Berlin, Germany

    Susanne M. Krug & Michael Fromm

  4. Leibniz-Institute of Molecular Pharmacology (FMP), Berlin-Buch, Berlin, Germany

    Jimmi D. Cording, Jörg Piontek & Ingolf E. Blasig

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  1. Julie K. Westphal
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Correspondence to Otmar Huber.

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18_2010_313_MOESM1_ESM.tif

Suppl. Fig. 1: (A) Western blot analysis of the expression of the FLAG-tagged tricellulin constructs in stably transfected MDCK C11 cell clones. Please note that clone K5 expresses higher levels of the full-length tricellulin construct compared to clone K13. (B) Localization of the tricellulin constructs in the different clones by confocal immunofluorescence microscopy after PFA fixation. In clone K5 localization of Tric-FLAG3 is not restricted to tricellular contacts but extends into the bicellular tight junctions as previously reported [4]. (TIFF 17745 kb)

18_2010_313_MOESM2_ESM.tif

Suppl. Fig. 2: Confocal immunofluorescence images (XYZ scans) reveals co-localization of the tricellulin constructs with ZO-1 at tight junctions. Please note that the TricDC-FLAG3 construct is less efficiently translocated to the tight junctions. For the image presented here, a cell that shows relatively strong membrane staining of TricDC-FLAG3 was chosen. (TIFF 26130 kb)

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Westphal, J.K., Dörfel, M.J., Krug, S.M. et al. Tricellulin forms homomeric and heteromeric tight junctional complexes. Cell. Mol. Life Sci. 67, 2057–2068 (2010). https://doi.org/10.1007/s00018-010-0313-y

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  • DOI: https://doi.org/10.1007/s00018-010-0313-y

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