CUED Publications database

Structured illumination microscopy reveals focal adhesions are composed of linear subunits.

Hu, S and Tee, Y-H and Kabla, A and Zaidel-Bar, R and Bershadsky, A and Hersen, P (2015) Structured illumination microscopy reveals focal adhesions are composed of linear subunits. In: UNSPECIFIED pp. 235-245..

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Abstract

The ability to mechanically interact with the extracellular matrix is a fundamental feature of adherent eukaryotic cells. Cell-matrix adhesion in many cell types is mediated by protein complexes called focal adhesions (FAs). Recent progress in super resolution microscopy revealed FAs possess an internal organization, yet such methods do not enable observation of the formation and dynamics of their internal structure in living cells. Here, we combine structured illumination microscopy (SIM) with total internal reflection fluorescence microscopy (TIRF) to show that the proteins inside FA patches are distributed along elongated subunits, typically 300 ± 100 nm wide, separated by 400 ± 100 nm, and individually connected to actin cables. We further show that the formation and dynamics of these linear subunits are intimately linked to radial actin fiber formation and actomyosin contractility. We found FA growth to be the result of nucleation of new linear subunits and their coordinated elongation. Taken together, this study reveals that the basic units of mature focal adhesion are 300-nm-wide elongated, dynamic structures. We anticipate this ultrastructure to be relevant to investigation of the function of FAs and their behavior in response to mechanical stress.

Item Type: Conference or Workshop Item (UNSPECIFIED)
Uncontrolled Keywords: cell mechanics cytoskeleton focal adhesion mechanobiology paxillin structured illumination microscopy Actin Cytoskeleton Actins Animals Cell Adhesion Cell Line Cytoskeleton Extracellular Matrix Fibroblasts Fibronectins Focal Adhesions Green Fluorescent Proteins Image Processing, Computer-Assisted Luminescent Proteins Microscopy, Fluorescence Paxillin Phalloidine Rats Stress Fibers Stress, Mechanical Vinculin
Subjects: UNSPECIFIED
Divisions: Div C > Biomechanics
Depositing User: Cron Job
Date Deposited: 17 Jul 2017 19:40
Last Modified: 24 Aug 2017 01:28
DOI: