title: ZO-1 interactions with F-actin and occludin direct epithelial polarization and single lumen specification in 3D culture.
abstract: Epithelia within tubular organs form and expand lumens. Failure of these processes can result in serious developmental anomalies. Although tight junction assembly is crucial to epithelial polarization, the contribution of specific tight junction proteins to lumenogenesis is undefined. Here, we show that ZO-1 (also known as TJP1) is necessary for the formation of single lumens. Epithelia lacking this tight junction scaffolding protein form cysts with multiple lumens and are defective in the earliest phases of polarization, both in two and three dimensions. Expression of ZO-1 domain-deletion mutants demonstrated that the actin-binding region and U5-GuK domain are crucial to single lumen development. For actin-binding region, but not U5-GuK domain, mutants, this could be overcome by strong polarization cues from the extracellular matrix. Analysis of the U5-GuK binding partners shroom2, α-catenin and occludin showed that only occludin deletion led to multi-lumen cysts. Like ZO-1-deficiency, occludin deletion led to mitotic spindle orientation defects. Single lumen formation required the occludin OCEL domain, which binds to ZO-1. We conclude that ZO-1-occludin interactions regulate multiple phases of epithelial polarization by providing cell-intrinsic signals that are required for single lumen formation.
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA.
authors: Matthew A
authors: MA
authors: Odenwald
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Wangsun
authors: W
authors: Choi
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Aaron
authors: A
authors: Buckley
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Nitesh
authors: N
authors: Shashikanth
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA.|Department of Pathology, NorthShore University Health System, Evanston, IL 60201, USA.
authors: Nora E
authors: NE
authors: Joseph
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA.
authors: Yitang
authors: Y
authors: Wang
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA.
authors: Michael H
authors: MH
authors: Warren
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA.
authors: Mary M
authors: MM
authors: Buschmann
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Roman
authors: R
authors: Pavlyuk
authors: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors: Jeffrey
authors: J
authors: Hildebrand
authors: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA.
authors: Ben
authors: B
authors: Margolis
authors: Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, NC 27599, USA.
authors: Alan S
authors: AS
authors: Fanning
authors: Department of Pathology, The University of Chicago, Chicago, IL 60637, USA jrturner@bwh.harvard.edu.|Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.|Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Jerrold R
authors: 0000-0003-0627-9455
authors: JR
authors: Turner
mesh_terms: D000199:Actins / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D016764:Cell Polarity; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D008938:Mitosis; D009024:Morphogenesis; D062793:Occludin / Q000378:metabolism*; D010641:Phenotype; D011485:Protein Binding; D021381:Protein Transport; D019108:Tight Junctions / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000737:chemistry / Q000378:metabolism*; D051178:alpha Catenin / Q000378:metabolism
chemical_list: D000199:Actins; D062793:Occludin; D062826:Zonula Occludens-1 Protein; D051178:alpha Catenin
keywords: Cell polarity; Epithelial Cells; Lumen Formation; Morphogenesis; Tight junctions