Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior

PLoS One. 2017 Apr 7;12(4):e0175324. doi: 10.1371/journal.pone.0175324. eCollection 2017.

Abstract

The extracellular matrix (ECM) is a major regulator of cell behavior. Recent studies have indicated the importance of the physical properties of the ECM, including its stiffness, for cell migration and differentiation. Using actomyosin-generated forces, cells pull the ECM and sense stiffness via cell-ECM adhesion structures called focal adhesions (FAs). Vinculin, an actin-binding FA protein, has emerged as a major player in FA-mediated mechanotransduction. Although vinculin is important for sensing ECM stiffness, the role of vinculin binding to actin in the ECM stiffness-mediated regulation of vinculin behavior remains unknown. Here, we show that an actin binding-deficient mutation disrupts the ECM stiffness-dependent regulation of CSB (cytoskeleton stabilization buffer) resistance and the stable localization of vinculin. These results suggest that the vinculin-actin interaction participates in FA-mediated mechanotransduction.

MeSH terms

  • Actin Cytoskeleton / metabolism*
  • Animals
  • Cells, Cultured
  • Extracellular Matrix / metabolism
  • Focal Adhesions
  • Mice
  • Protein Binding
  • Subcellular Fractions / metabolism
  • Vinculin / genetics
  • Vinculin / metabolism*

Substances

  • Vinculin

Grants and funding

This work was supported in part by The Naito Foundation(NK), a Grant-in-Aid for Scientific Research (B) (JSPS KAKENHI Grant Number 24380185)(NK), a Grant-in-aid for Exploratory Research (JSPS KAKENHI Grant Number 26660291, 16K15090)(NK) from The Japan Society for the Promotion of Science (https://www.jsps.go.jp/english/index.html), Grants-in-Aid for Scientific Research (S) (25221203)(KU) and a Grant-in-Aid for Scientific Research on Innovative Areas (MEXT KAKENHI Grant Number 26112707)(NK) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (http://www.mext.go.jp/en/), and the Advanced Research and Development Programs for Medical Innovation (NK) from Japan Agency for Medical Research and development (http://www.amed.go.jp/en/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.