title: Vimentin Levels and Serine 71 Phosphorylation in the Control of Cell-Matrix Adhesions, Migration Speed, and Shape of Transformed Human Fibroblasts.
abstract: Metastasizing tumor cells show increased expression of the intermediate filament (IF) protein vimentin, which has been used to diagnose invasive tumors for decades. Recent observations indicate that vimentin is not only a passive marker for carcinoma, but may also induce tumor cell invasion. To clarify how vimentin IFs control cell adhesions and migration, we analyzed the nanoscale (30-50 nm) spatial organization of vimentin IFs and cell-matrix adhesions in metastatic fibroblast cells, using three-color stimulated emission depletion (STED) microscopy. We also studied whether wild-type and phospho-deficient or -mimicking mutants of vimentin changed the size and lifetime of focal adhesions (FAs), cell shape, and cell migration, using live-cell total internal reflection imaging and confocal microscopy. We observed that vimentin exists in fragments of different lengths. Short fragments were mostly the size of a unit-length filament and were mainly localized close to small cell-matrix adhesions. Long vimentin filaments were found in the proximity of large FAs. Vimentin expression in these cells caused a reduction in FAs size and an elongated cell shape, but did not affect FA lifetime, or the speed or directionality of cell migration. Expression of a phospho-mimicking mutant (S71D) of vimentin increased the speed of cell migration. Taken together, our results suggest that in highly migratory, transformed mesenchymal cells, vimentin levels control the cell shape and FA size, but not cell migration, which instead is linked to the phosphorylation status of S71 vimentin. These observations are consistent with the possibility that not only levels, but also the assembly status of vimentin control cell migration.
authors.affiliation: Department of Physics, University of the Saarland, 66123 Saarbrücken, Germany. emmanuel.terriac@physik.uni-saarland.de.|INM - Leibniz Institute for New Materials, 66123 Saarbrücken, Germany. emmanuel.terriac@physik.uni-saarland.de.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Terriac
authors.affiliation: Department of Applied Physics, KTH Royal Institute of Technology, 100 44 Stockholm, Sweden. giovanna.coceano@scilifelab.se.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Coceano
authors.affiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, 171 65 Stockholm, Sweden. Mavajian@kth.se.|Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden. Mavajian@kth.se.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Mavajian
authors.affiliation: KIST Europe, 66123 Saarbrücken, Germany. t.hageman@kist-europe.de.
authors.forename: Tijmen A G
authors.initials: TA
authors.lastname: Hageman
authors.affiliation: Department of Physics, University of the Saarland, 66123 Saarbrücken, Germany. andreas.f.christ@gmail.com.
authors.forename: Andreas F
authors.initials: AF
authors.lastname: Christ
authors.affiliation: Department of Applied Physics, KTH Royal Institute of Technology, 100 44 Stockholm, Sweden. ilaria.testa@scilifelab.se.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Testa
authors.affiliation: Department of Physics, University of the Saarland, 66123 Saarbrücken, Germany. f.lautenschlaeger@physik.uni-saarland.de.|INM - Leibniz Institute for New Materials, 66123 Saarbrücken, Germany. f.lautenschlaeger@physik.uni-saarland.de.
authors.forename: Franziska
authors.initials: F
authors.lastname: Lautenschläger
authors.affiliation: Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, 171 65 Stockholm, Sweden. Annica.Gad@ki.se.|Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77 Stockholm, Sweden. Annica.Gad@ki.se.
authors.forename: Annica K B
authors.initials: AK
authors.lastname: Gad
keywords: cell migration; focal adhesions; stimulated emission depletion (STED) microscopy; total internal reflection (TIRF) microscopy; vimentin