The coordinated interplay of cytoskeletal networks critically determines tissue biomechanics and structural integrity. Here, we show that plectin, a major intermediate filament-based cytolinker protein, orchestrates cortical cytoskeletal networks in epithelial sheets to support intercellular junctions. By combining CRISPR/Cas9-based gene editing and pharmacological inhibition, we demonstrate that in an F-actin-dependent context, plectin is essential for the formation of the circumferential keratin rim, organization of radial keratin spokes, and desmosomal patterning. In the absence of plectin-mediated cytoskeletal cross-linking, the aberrant keratin-desmosome (DSM)-network feeds back to the actin cytoskeleton, which results in elevated actomyosin contractility. Also, by complementing a predictive mechanical model with Förster resonance energy transfer-based tension sensors, we provide evidence that in the absence of cytoskeletal cross-linking, major intercellular junctions (adherens junctions and DSMs) are under intrinsically generated tensile stress. Defective cytoarchitecture and tensional disequilibrium result in reduced intercellular cohesion, associated with general destabilization of plectin-deficient sheets upon mechanical stress.

Department of Analytical Chemistry University of Vienna Vienna Austria

Department of Biochemistry and Cell Biology Max Perutz Labs University of Vienna Vienna Austria

Department of Physics University of Erlangen Nuremberg Erlangen Germany

Department of Physiology Faculty of Science Charles University Prague Czech Republic

Department of Quantitative Cell Biology Institute of Molecular Cell Biology University of Münster Münster Germany

Laboratory of Integrative Biology Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

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J Cell Biol. 2022 Mar 7;221(3):e202201054. doi: 10.1083/jcb.202201054. PubMed

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Mechanics of cell sheets: plectin as an integrator of cytoskeletal networks