Regulation of podocalyxin trafficking by Rab small GTPases in 2D and 3D epithelial cell cultures

Paulina S. Mrozowska, Mitsunori Fukuda

    Research output: Contribution to journalArticlepeer-review

    49 Citations (Scopus)

    Abstract

    MDCK II cells, a widely used model of polarized epithelia, develop into different structures depending on culture conditions: two-dimensional (2D) monolayers when grown on synthetic supports or three-dimensional (3D) cysts when surrounded by an extracellular matrix. The establishment of epithelial polarity is accompanied by transcytosis of the apical marker podocalyxin from the outer plasma membrane to the newly formed apical domain, but its exact route and regulation remain poorly understood. Here, through comprehensive colocalization and knockdown screenings, we identified the Rab GTPases mediating podocalyxin transcytosis and showed that different sets of Rabs coordinate its transport during cell polarization in 2D and 3D structures. Moreover, we demonstrated that different Rab35 effectors regulate podocalyxin trafficking in 2D and 3D environments; trafficking is mediated by OCRL in 2D monolayers and ACAP2 in 3D cysts. Our results give substantial insight into regulation of the transcytosis of this apical marker and highlight differences between trafficking mechanisms in 2D and 3D cell cultures.

    Original languageEnglish
    Pages (from-to)355-369
    Number of pages15
    JournalJournal of Cell Biology
    Volume213
    Issue number3
    DOIs
    Publication statusPublished - 2016 May 9

    ASJC Scopus subject areas

    • Cell Biology

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