Abstract
The presence of tight junction protein zonula occludens 2 (ZO-2) at the nucleus inhibits the transcription of genes regulated by TEAD transcription factor. Here, we analyzed whether the movement of ZO-2 into the nucleus modulates the nuclear concentration of TEAD. In sparse cultures of ZO-2 knockdown Madin-Darby canine kidney cells, nuclear TEAD was diminished, as in parental cells transfected with a ZO-2 construct without nuclear localization signals, indicating that ZO-2 facilitates the entry of TEAD into the nucleus. Inhibition of nPKCδ in parental cells triggers the interaction between ZO-2 and TEAD at the cytoplasm and facilitates TEAD/ZO-2 complex nuclear importation. Using proximity ligation, immunoprecipitation, and pull-down assays, TEAD/ZO-2 interaction was confirmed. Nuclear TEAD is phosphorylated, and its exit in parental cells is enhanced by activation of a ZO-2 nuclear exportation signal by nPKCε, while the nuclear accumulation of ZO-2 triggered by the mutation of ZO-2 nuclear export signals induces no change in TEAD nuclear concentration. In summary, our results indicate that the movements of ZO-2 in and out of the nucleus modulate the intracellular traffic of TEAD through a process regulated by nPKCδ and ε and provide a novel role of ZO-2 as a nuclear translocator of TEAD.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line
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Cell Nucleus / metabolism*
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Dogs
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Epithelial Cells / metabolism*
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HEK293 Cells
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Humans
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Nuclear Localization Signals
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Phosphorylation
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Protein Binding
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Protein Kinase C-epsilon / metabolism
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Protein Processing, Post-Translational
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Protein Transport
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Rats
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Signal Transduction
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TEA Domain Transcription Factors / metabolism*
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Zonula Occludens-2 Protein / metabolism*
Substances
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Nuclear Localization Signals
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TEA Domain Transcription Factors
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Zonula Occludens-2 Protein
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Protein Kinase C-epsilon