The Keratin Cortex Stabilizes Cells at High Strains

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract The eukaryotic cytoskeleton consists of three filament types: actin filaments, microtubules and intermediate filaments (IFs). IF proteins are expressed in a cell-type specific manner, and keratins are found in epithelial cells. In certain cell types, keratin forms a layer close to the membrane which may be referred to as an “IF-cortex”. It is hypothesized that this IF-cortex arranges with radial bundles in a “rim-and-spokes” structure in epithelia. Based on this hypothesis, IFs and actin filaments might add complementary mechanical properties to the cortex. It was previously shown that single IFs in vitro remain undamaged at high strains and display a non-linear stretching behavior. We now ask the question of whether this unique force-extension behavior of single IFs is also relevant in the context of a filament network within a cell. We show that keratin-deficient (KO) MDCK II cells readily form 2D cell layers and 3D cysts and withstand high equibiaxial strains. High-resolution imaging using STED microscopy reveals altered actin cortex structures in KO cells, presumably in response to the missing keratin. We investigate the influence of the equibiaxial strain on the viscoelastic properties of wild-type (WT) and KO cells using atomic force microscopy. We find that the KO cells exhibit a higher pre-stress than the WT cells, likely due to the change of the cortical structure. Interestingly, both the pre-stress and the fluidity of the KO cells are altered already at intermediate strains, whereas the WT cells show a response only at high strain. Similarly, the KO cysts are stretched more easily at low strains than the WT cysts during injection experiments. The compressibility modulus is analyzed in a spatially resolved manner and we find this modulus to be increased at the cell rim, compared to the inside region, due to the geometry of the cell layer. Our results indicate that KO cells compensate for the missing keratin, but are nevertheless very sensitive to external strain, whereas the intricate interplay between the actin and keratin cortices in WT cells preserves the mechanical state and cell stability.

Authors 5

1. Nicole Schwarz Aachen Institute of Molecular and Cellular Anatomy (MOCA)

   RWTH Aachen University

   Affiliation as printed

   Institute of Molecular and Cellular Anatomy, RWTH Aachen University, Wendlingweg 2, 52074 Aachen, Germany;

2. Amaury Perez-Tirado

   University of Göttingen

   Affiliation as printed

   Institute of Physical Chemistry, University of Gottingen, Tammannstrasse 6, 37077 Gottingen, Germany;

3. Anna V. Schepers

   University of Göttingen

   Affiliation as printed

   Institute for X-Ray Physics, University of Göttingen, Friedrich-Hund-Platz 1, 37077 Göottingen, Germany;

4. Andreas Janshoff

   Multiscale Bioimaging · Max Planck School Matter to Life · University of Göttingen

   Affiliation as printed

   Institute of Physical Chemistry, University of Gottingen, Tammannstrasse 6, 37077 Gottingen, Germany; Max Planck School ``Matter to Life''; Cluster of Excellence ``Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells'' (MBE

5. Sarah Köster corresponding

   University of Göttingen

   Affiliation as printed

   Institute for X-Ray Physics, University of Göttingen, Friedrich-Hund-Platz 1, 37077 Göottingen, Germany;