Endoplasmic spreading requires coalescence of vimentin intermediate filaments at force-bearing adhesions

Christopher D. Lynch, Andre M. Lazar, Thomas Iskratsch, Xian Zhang, Michael Sheetz

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

For cells to develop long-range forces and carry materials to the periphery, the microtubule and organelle-rich region at the center of the cell - the endoplasm - needs to extend to near the cell edge. Depletion of the actin cross-linking protein filamin A (FlnA) causes a collapse of the endoplasm into a sphere around the nucleus of fibroblasts and disruption of matrix adhesions, indicating that FlnA is involved in endoplasmic spreading and adhesion growth. Here, we report that treatment with the calpain inhibitor N-[N-(N-acetyl-L- leucyl)-L-leucyl]-L-norleucine (ALLN) restores endoplasmic spreading as well as focal adhesion (FA) growth on fibronectin-coated surfaces in a Fln-depleted background. Addback of calpain-uncleavable talin, not full-length talin, achieves a similar effect in Fln-depleted cells and indicates a crucial role for talin in endoplasmic spreading. Because FA maturation involves the vimentin intermediate filament (vIF) network, we also examined the role of vIFs in endoplasmic spreading. Wild-type cells expressing a vimentin variant incapable of polymerization exhibit deficient endoplasmic spreading as well as defects in FA growth. ALLN treatment restores FA growth despite the lack of vIFs but does not restore endoplasmic spreading, implying that vIFs are essential for endoplasm spreading. Consistent with that hypothesis, vIFs are always displaced from adhesions when the endoplasm does not spread. In Fln-depleted cells, vIFs extend beyond adhesions, nearly to the cell edge. Finally, inhibiting myosin II-mediated contraction blocks endoplasmic spreading and adhesion growth. Thus we propose a model in which myosin II-mediated forces and coalescence of vIFs at mature FAs are required for endoplasmic spreading.

Original languageEnglish (US)
Pages (from-to)21-30
Number of pages10
JournalMolecular Biology of the Cell
Volume24
Issue number1
DOIs
StatePublished - Jan 1 2013
Externally publishedYes

Fingerprint

Intermediate Filaments
Vimentin
Focal Adhesions
Talin
Filamins
Myosin Type II
Growth
Calpain
Fibronectins
Microtubules
Polymerization
Organelles
Actins
Fibroblasts

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Endoplasmic spreading requires coalescence of vimentin intermediate filaments at force-bearing adhesions. / Lynch, Christopher D.; Lazar, Andre M.; Iskratsch, Thomas; Zhang, Xian; Sheetz, Michael.

In: Molecular Biology of the Cell, Vol. 24, No. 1, 01.01.2013, p. 21-30.

Research output: Contribution to journalArticle

Lynch, Christopher D. ; Lazar, Andre M. ; Iskratsch, Thomas ; Zhang, Xian ; Sheetz, Michael. / Endoplasmic spreading requires coalescence of vimentin intermediate filaments at force-bearing adhesions. In: Molecular Biology of the Cell. 2013 ; Vol. 24, No. 1. pp. 21-30.
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