Rho-associated kinase-dependent contraction of stress fibres and the organization of focal adhesions

Katoh, Kazuo; Kano, Yumiko; Noda, Yasuko

doi:10.1098/rsif.2010.0419

Cited by 100 publications

(95 citation statements)

References 59 publications

(114 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MLCK could trigger MLC phosphorylation to enhance the actin-myosin interaction, which in turn increased the cell tension [50]. Without that, cells may maintain the cytoskeleton structure, but lose the cellular tension [51]. As the result, hMSCs with MLCK inhibited was unable to commit to myogenic differentiation in our study.…”

Section: Discussionmentioning

confidence: 64%

Insights into the Role of Focal Adhesion Modulation in Myogenic Differentiation of Human Mesenchymal Stem Cells

Lui

Xiong

et al. 2013

Stem Cells and Development

View full text Add to dashboard Cite

We report the establishment of a novel platform to induce myogenic differentiation of human mesenchymal stem cells (hMSCs) via focal adhesion (FA) modulation, giving insights into the role of FA on stem cell differentiation. Micropatterning of collagen type I on a polyacrylamide gel with a stiffness of 10.2 kPa efficiently modulated elongated FA. This elongated FA profile preferentially recruited the b 3 integrin cluster and induced specific myogenic differentiation at both transcription and translation levels with expression of myosin heavy chain and a-sarcomeric actin. This was initiated with elongation of FA complexes that triggered the RhoA downstream signaling toward a myogenic lineage commitment. This study also illustrates how one could partially control myogenic differentiation outcomes of similar-shaped hMSCs by modulating FA morphology and distribution. This technology increases our toolkit choice for controlled differentiation in muscle engineering.

show abstract

Section: Discussionmentioning

confidence: 64%

Insights into the Role of Focal Adhesion Modulation in Myogenic Differentiation of Human Mesenchymal Stem Cells

Lui

Xiong

et al. 2013

Stem Cells and Development

View full text Add to dashboard Cite

show abstract

“…The retraction dynamics yield quantitative information about the viscoelastic properties of the fiber, and the resulting change in cell shape lends insight into the contribution of the targeted SF to cell shape stability. We have previously used this approach to demonstrate that the mechanical properties of a SF depend strongly on whether the fiber is located at the cell center or periphery (Tanner et al, 2010), and that this is related to the differential control of these two SF populations by the myosin activators Rho-associated kinase (ROCK) and myosin light chain kinase (MLCK), respectively (Katoh et al, 2011;Katoh et al, 2007;Totsukawa et al, 2004). Moreover, disruption of peripheral SFs often triggers whole-cell adhesive disassembly and contraction, which has led us to speculate that central and peripheral SFs distribute their tensile loads differently across the cell-ECM interface and that this in turn gives rise to their differential contributions to cell shape stability.…”

Section: Introductionmentioning

confidence: 99%

Vinculin tension distributions of individual stress fibers within cell-matrix adhesions

Chang

Kumar

2013

Journal of Cell Science

View full text Add to dashboard Cite

SummaryActomyosin stress fibers (SFs) enable cells to exert traction on planar extracellular matrices (ECMs) by tensing focal adhesions (FAs) at the cell-ECM interface. Although it is widely appreciated that the spatial and temporal distribution of these tensile forces play key roles in polarity, motility, fate choice, and other defining cell behaviors, virtually nothing is known about how an individual SF quantitatively contributes to tensile loads borne by specific molecules within associated FAs. We address this key open question by using femtosecond laser ablation to sever single SFs in cells while tracking tension across vinculin using a molecular optical sensor. We show that disruption of a single SF reduces tension across vinculin in FAs located throughout the cell, with enriched vinculin tension reduction in FAs oriented parallel to the targeted SF. Remarkably, however, some subpopulations of FAs exhibit enhanced vinculin tension upon SF irradiation and undergo dramatic, unexpected transitions between tension-enhanced and tension-reduced states. These changes depend strongly on the location of the severed SF, consistent with our earlier finding that different SF pools are regulated by distinct myosin activators. We critically discuss the extent to which these measurements can be interpreted in terms of whole-FA tension and traction and propose a model that relates SF tension to adhesive loads and cell shape stability. These studies represent the most direct and highresolution intracellular measurements of SF contributions to tension on specific FA proteins to date and offer a new paradigm for investigating regulation of adhesive complexes by cytoskeletal force.

show abstract

“…During contraction of stress fibers, myosin mediates sliding of antiparallel actin filaments and plays a key role in stress fiber stability and cell motility (Hotulainen and Lappalainen, 2006). Notably, phosphorylation of MLC2 has been demonstrated to be a vital motor involved in motility (Chaturvedi et al, 2011;Katoh et al, 2011). Therefore, we investigated the expression and phosphorylation of MLC2 in our NDRG1 overexpression and knockdown models.…”

Section: Ndrg1 Inhibits F-actin Polymerization and Stressmentioning

confidence: 99%

“…It is reported that a key factor in stress fiber formation is the phosphorylation of myosin light chain 2 (MLC2) at Thr18/ Ser19 (Katoh et al, 2011). Cells have a complicated signaling network modulating MLC2 phosphorylation, among which the Rho-associated, coiled-coil-containing protein kinase 1 (ROCK1) pathway is one of the most crucial mechanisms in stimulating cross-linking of actin by myosin, leading to enhanced cancer cell contractility (Chaturvedi et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Targeting the Metastasis Suppressor, NDRG1, Using Novel Iron Chelators: Regulation of Stress Fiber-Mediated Tumor Cell Migration via Modulation of the ROCK1/pMLC2 Signaling Pathway

et al. 2012

View full text Add to dashboard Cite

The iron-regulated metastasis suppressor, N-myc downstreamregulated gene 1 (NDRG1), is up-regulated by cellular iron depletion mediated by iron chelators and can inhibit cancer cell migration. However, the mechanism of how NDRG1 achieves this effect remains unclear. In this study, we implemented established and newly constructed NDRG1 overexpression and knockdown models using the DU145, HT29, and HCT116 cancer cell lines to investigate the molecular basis by which NDRG1 exerts its inhibitory effect on cell migration. Using these models, we demonstrated that NDRG1 overexpression inhibits cell migration by preventing actin-filament polymerization, stress fiber assembly and formation. In contrast, NDRG1 knockdown had the opposite effect. Moreover, we identified that NDRG1 inhibited an important regulatory pathway mediated by the Rho-associated, coiled-coil containing protein kinase 1 (ROCK1)/phosphorylated myosin light chain 2 (pMLC2) pathway that modulates stress fiber assembly. The phosphorylation of MLC2 is a key process in inducing stress fiber contraction, and this was shown to be markedly decreased or increased by NDRG1 overexpression or knockdown, respectively. The mechanism involved in the inhibition of MLC2 phosphorylation by NDRG1 was mediated by a significant (P , 0.001) decrease in ROCK1 expression that is a key kinase involved in MLC2 phosphorylation. Considering that NDRG1 is up-regulated after cellular iron depletion, novel thiosemicarbazone iron chelators (e.g., di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone) were demonstrated to inhibit ROCK1/pMLC2-modulated actinfilament polymerization, stress fiber assembly, and formation via a mechanism involving NDRG1. These results highlight the role of the ROCK1/pMLC2 pathway in the NDRG1-mediated antimetastatic signaling network and the therapeutic potential of iron chelators at inhibiting metastasis.

show abstract

Rho-associated kinase-dependent contraction of stress fibres and the organization of focal adhesions

Cited by 100 publications

References 59 publications

Insights into the Role of Focal Adhesion Modulation in Myogenic Differentiation of Human Mesenchymal Stem Cells

Insights into the Role of Focal Adhesion Modulation in Myogenic Differentiation of Human Mesenchymal Stem Cells

Vinculin tension distributions of individual stress fibers within cell-matrix adhesions

Targeting the Metastasis Suppressor, NDRG1, Using Novel Iron Chelators: Regulation of Stress Fiber-Mediated Tumor Cell Migration via Modulation of the ROCK1/pMLC2 Signaling Pathway

Contact Info

Product

Resources

About