title: Tropomyosin isoforms support actomyosin biogenesis to generate contractile tension at the epithelial zonula adherens.
abstract: Epithelial cells generate contractile forces at their cell-cell contacts. These are concentrated at the specialized apical junction of the zonula adherens (ZA), where a ring of stabilized E-cadherin lies adjacent to prominent actomyosin bundles. Coupling of adhesion and actomyosin contractility yields tension in the junction. The biogenesis of junctional contractility requires actin assembly at the ZA as well as the recruitment of nonmuscle myosin II, but the molecular regulators of these processes are not yet fully understood. We now report a role for tropomyosins 5NM1 (Tm5NM1) and 5NM2 (Tm5NM2) in their generation. Both these tropomyosin isoforms were found at the ZA and their depletion by RNAi or pharmacological inhibition reduced both F-actin and myosin II content at the junction. Photoactivation analysis revealed that the loss of F-actin was attributable to a decrease in filament stability. These changes were accompanied by a decrease in E-cadherin content at junctions. Ultimately, both long-term depletion of Tm5NM1/2 and acute inhibition with drugs caused junctional tension to be reduced. Thus these tropomyosin isoforms are novel contributors to junctional contractility and integrity.
authors.affiliation: Division of Cell Biology and Molecular Medicine, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Caldwell
authors.forename: Christine
authors.initials: C
authors.lastname: Lucas
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Kee
authors.forename: Katharina
authors.initials: K
authors.lastname: Gaus
authors.forename: Peter W
authors.initials: PW
authors.lastname: Gunning
authors.forename: Edna C
authors.initials: EC
authors.lastname: Hardeman
authors.forename: Alpha S
authors.initials: AS
authors.lastname: Yap
authors.forename: Guillermo A
authors.initials: GA
authors.lastname: Gomez
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000205:Actomyosin / Q000378:metabolism*; D022005:Adherens Junctions / Q000378:metabolism*; D000818:Animals; D018938:Caco-2 Cells; D002450:Cell Communication / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D020033:Protein Isoforms / Q000378:metabolism; D034741:RNA, Small Interfering; D014162:Transfection; D014335:Tropomyosin / Q000378:metabolism*
chemical_list: D020033:Protein Isoforms; D034741:RNA, Small Interfering; D014335:Tropomyosin; D000205:Actomyosin
keywords: E-cadherin; actomyosin; contractility; tropomyosin; zonula adherens