The scaffolding protein ZO-1 coordinates actomyosin and epithelial apical specializations in vitro and in vivo

J Biol Chem. 2018 Nov 9;293(45):17317-17335. doi: 10.1074/jbc.RA118.003908. Epub 2018 Sep 21.


Polarized epithelia assemble into sheets that compartmentalize organs and generate tissue barriers by integrating apical surfaces into a single, unified structure. This tissue organization is shared across organs, species, and developmental stages. The processes that regulate development and maintenance of apical epithelial surfaces are, however, undefined. Here, using an intestinal epithelial-specific knockout (KO) mouse and cultured epithelial cells, we show that the tight junction scaffolding protein zonula occludens-1 (ZO-1) is essential for development of unified apical surfaces in vivo and in vitro We found that U5 and GuK domains of ZO-1 are necessary for proper apical surface assembly, including organization of microvilli and cortical F-actin; however, direct interactions with F-actin through the ZO-1 actin-binding region (ABR) are not required. ZO-1 lacking the PDZ1 domain, which binds claudins, rescued apical structure in ZO-1-deficient epithelia, but not in cells lacking both ZO-1 and ZO-2, suggesting that heterodimerization with ZO-2 restores PDZ1-dependent ZO-1 interactions that are vital to apical surface organization. Pharmacologic F-actin disruption, myosin II motor inhibition, or dynamin inactivation restored apical epithelial structure in vitro and in vivo, indicating that ZO-1 directs epithelial organization by regulating actomyosin contraction and membrane traffic. We conclude that multiple ZO-1-mediated interactions contribute to coordination of epithelial actomyosin function and genesis of unified apical surfaces.

Keywords: PDZ domain; actin; apical specialization; brush border; cell adhesion; cell polarity; cell surface; community border; cytoskeleton; intestinal epithelium; microvilli; myosin; tight junction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actins / genetics
  • Actins / metabolism
  • Actomyosin / genetics
  • Actomyosin / metabolism*
  • Animals
  • Biological Transport, Active / physiology
  • Cell Membrane / genetics
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Dynamins / genetics
  • Dynamins / metabolism
  • Epithelial Cells / metabolism*
  • Epithelial Cells / ultrastructure
  • Intestinal Mucosa / metabolism*
  • Intestinal Mucosa / ultrastructure
  • Mice
  • Mice, Knockout
  • Microvilli / genetics
  • Microvilli / metabolism*
  • Microvilli / ultrastructure
  • Myosin Type II / genetics
  • Myosin Type II / metabolism
  • Protein Multimerization / physiology
  • Zonula Occludens-1 Protein / genetics
  • Zonula Occludens-1 Protein / metabolism*
  • Zonula Occludens-2 Protein / genetics
  • Zonula Occludens-2 Protein / metabolism


  • Actins
  • Tjp1 protein, mouse
  • Tjp2 protein, mouse
  • Zonula Occludens-1 Protein
  • Zonula Occludens-2 Protein
  • Actomyosin
  • Myosin Type II
  • Dynamins