Specialized actin nanoscale layers control focal adhesion turnover

Abstract

Focal adhesions (FAs) connect inner workings of the cell to the extracellular matrix to control cell adhesion, migration, and mechanosensing. Previous studies demonstrated that FAs contain three vertical layers, which connect extracellular matrix to the cytoskeleton. However, cellular processes rely on precisely-regulated FA turnover, but the molecular machineries that control FA assembly and disassembly have remained elusive. By using super-resolution iPALM microscopy, we identified two unprecedented nanoscale layers within FAs, specified by actin filaments bound to tropomyosin isoforms Tpm1.6 and Tpm3.2. The Tpm1.6-actin filaments beneath the previously identified actin-regulatory layer are critical for adhesion maturation and controlled cell motility, whereas the Tpm3.2-actin filament layer towards the bottom of FA facilitates adhesion disassembly. Mechanistically, Tpm3.2 stabilizes KANK-family proteins at adhesions, and hence targets microtubule plus-ends to FAs to catalyse their disassembly. Loss of Tpm3.2 leads to disorganized microtubule network, abnormally stable FAs, and defects in tail retraction during cell migration. Thus, FAs are composed of at least three distinct actin filament layers, each having specific roles in coupling of adhesion to the cytoskeleton, or in controlling adhesion dynamics. In a broader context, these findings demonstrate how distinct actin filament populations can co-exist and perform specific functions within a defined cellular compartment.

Functional redundancy and formin-independent localization of tropomyosin isoforms inSaccharomyces cerevisiae

“…Structural analysis using cryoEM has now revealed that Tpm isoforms may exert differential functions via different modes of binding and controlling accessibility of ABPs like myosin and cofilin to F-actin 29,30 . How Tpm isoforms have been fine-tuned to perform distinct spatial sorting and functions during evolution remains an active area of research and their crosstalk during recruitment and maturation within the same actin structure is just beginning to be understood 21,31 .…”