Tight junction protein ZO-2 modulates the nuclear accumulation of transcription factor TEAD

• Published in: Molecular biology of the cell Vol. 32; no. 15; pp. 1347 - 1358
• Main Authors: Gallego-Gutiérrez, Helios, González-González, Laura, Ramírez-Martínez, Leticia, López-Bayghen, Esther, González-Mariscal, Lorenza
• Format: Journal Article
• Language: English
• Published: United States The American Society for Cell Biology 15.07.2021
• Subjects: Animals; Cell Line; Cell Nucleus - metabolism; Dogs; Epithelial Cells - metabolism; HEK293 Cells; Humans; Nuclear Localization Signals; Phosphorylation; Protein Binding; Protein Kinase C-epsilon - metabolism; Protein Processing, Post-Translational; Protein Transport; Rats; Signal Transduction; TEA Domain Transcription Factors - metabolism; Zonula Occludens-2 Protein - metabolism
• ISSN: 1059-1524; 1939-4586
• DOI: 10.1091/mbc.E20-07-0470

The presence of tight junction protein onula ccludens 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.