The Role of ZO-2 in Modulating JAM-A and γ-Actin Junctional Recruitment, Apical Membrane and Tight Junction Tension, and Cell Response to Substrate Stiffness and Topography

Diana Cristina Pinto-Dueñas, Christian Hernández-Guzmán, Patrick Matthew Marsch, Anand Sunil Wadurkar, Dolores Martín-Tapia, Lourdes Alarcón, Genaro Vázquez-Victorio, Juan Vicente Méndez-Méndez, José Jorge Chanona-Pérez, Shikha Nangia, Lorenza González-Mariscal

Research output: Contribution to journalArticlepeer-review

Abstract

This work analyzes the role of the tight junction (TJ) protein ZO-2 on mechanosensation. We found that the lack of ZO-2 reduced apical membrane rigidity measured with atomic force microscopy, inhibited the association of γ-actin and JAM-A to the cell border, and instead facilitated p114RhoGEF and afadin accumulation at the junction, leading to an enhanced mechanical tension at the TJ measured by FRET, with a ZO-1 tension probe, and increased tricellular TJ tension. Simultaneously, adherens junction tension measured with an E-cadherin probe was unaltered. The stability of JAM-A and ZO-2 binding was assessed by a collaborative in silico study. The absence of ZO-2 also impacted the cell response to the substrate, as monolayers plated in 20 kPa hydrogels developed holes not seen in parental cultures and displayed a retarded elongation and formation of cell aggregates. The absence of ZO-2 was sufficient to induce YAP and Snail nuclear accumulation in cells cultured over glass, but when ZO-2 KD cells were plated in nanostructured ridge arrays, they displayed an increased abundance of nuclear Snail and conspicuous internalization of claudin-4. These results indicate that the absence of ZO-2 also impairs the response of cells to substrate stiffness and exacerbates transformation triggered by substrate topography.

Original languageEnglish (US)
Article number2453
JournalInternational journal of molecular sciences
Volume25
Issue number5
DOIs
StatePublished - Mar 2024

Keywords

  • JAM-A
  • ZO-2
  • afadin
  • p114RhoGEF
  • tension
  • tight junction
  • γ-actin

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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