scholarly journals Focal adhesion-generated cues in extracellular matrix regulate cell migration by local induction of clathrin-coated plaques

2018 ◽  
Author(s):  
Delia Bucher ◽  
Markus Mukenhirn ◽  
Kem A. Sochacki ◽  
Veronika Saharuka ◽  
Christian Huck ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Focal Adhesions ◽  
Plaque Formation ◽  
Cell Contact ◽  
Coated Pits ◽  
Independent Manner ◽  
A Cell ◽  
And Function ◽  
Topographical Cues

AbstractClathrin is a unique scaffold protein, which forms polyhedral lattices with flat and curved morphology. The function of curved clathrin-coated pits in forming endocytic structures is well studied. On the contrary, the role of large flat clathrin arrays, called clathrin-coated plaques, remains ambiguous. Previous studies suggested an involvement of plaques in cell adhesion. However, the molecular origin leading to their formation and their precise functions remain to be determined. Here, we study the origin and function of clathrin-coated plaques during cell migration. We revealed that plaque formation is intimately linked to extracellular matrix (ECM) modification by focal adhesions (FAs). We show that in migrating cells, FAs digest the ECM creating extracellular topographical cues that dictate the future location of clathrin-coated plaques. We identify Eps15 and Eps15R as key regulators for the formation of clathrin-coated plaques at locally remodelled ECM sites. Using a genetic silencing approach to abrogate plaque formation and 3D-micropatterns to spatially control the location of clathrin-coated plaques, we could directly correlate cell migration directionality with the formation of clathrin-coated plaques and their ability to recognize extracellular topographical cues. We here define the molecular mechanism regulating the functional interplay between FAs and plaques and propose that clathrin-coated plaques act as regulators of cell migration promoting contact guidance-mediated collective migration in a cell-to-cell contact independent manner.

scholarly journals Mitochondria tether to Focal Adhesions during cell migration and regulate their size

2019 ◽  
Author(s):  
Redaet Daniel ◽  
Abebech Mengeta ◽  
Patricia Bilodeau ◽  
Jonathan M Lee
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Energy Production ◽  
Focal Adhesions ◽  
Leading Edge ◽  
Energy Requirements ◽  
Integrin Receptors ◽  
Cellular Energy ◽  
A Cell ◽  
Atp Generation

AbstractMitochondria are the key generators of ATP in a cell. Visually, they are highly dynamic organelles that undergo cellular fission and fusion events in response to changing cellular energy requirements. Mitochondria are now emerging as regulators of mammalian cell motility. Here we show that mitochondria infiltrate the leading edge of NIH3T3 fibroblasts during migration. At the leading edge, we find that mitochondria move to and tether to Focal Adhesions (FA). FA regulate cell migration by coupling the cytoskeleton to the Extracellular Matrix through integrin receptors. Importantly, we find that inhibition of mitochondrial ATP generation concomitantly inhibits FA size. This suggests that mitochondrial energy production regulates migration through FA control.

Syndesmos, a protein that interacts with the cytoplasmic domain of syndecan-4, mediates cell spreading and actin cytoskeletal organization

2000 ◽  
Vol 113 (2) ◽  
pp. 315-324 ◽  
Author(s):  
P.C. Baciu ◽  
S. Saoncella ◽  
S.H. Lee ◽  
F. Denhez ◽  
D. Leuthardt ◽  
...  
Keyword(s):  
Plasma Membranes ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Cytoplasmic Domain ◽  
Cellular Protein ◽  
Actin Stress Fiber ◽  
Independent Manner ◽  
Intracellular Proteins ◽  
Central Regions ◽  
A Cell

Syndecan-4 is a cell surface heparan sulfate proteoglycan which, in cooperation with integrins, transduces signals for the assembly of focal adhesions and actin stress fibers in cells plated on fibronectin. The regulation of these cellular events is proposed to occur, in part, through the interaction of the cytoplasmic domains of these transmembrane receptors with intracellular proteins. To identify potential intracellular proteins that interact with the cytoplasmic domain of syndecan-4, we carried out a yeast two-hybrid screen in which the cytoplasmic domain of syndecan-4 was used as bait. As a result of this screen, we have identified a novel cellular protein that interacts with the cytoplasmic domain of syndecan-4 but not with those of the other three syndecan family members. The interaction involves both the membrane proximal and variable central regions of the cytoplasmic domain. We have named this cDNA and encoded protein syndesmos. Syndesmos is ubiquitously expressed and can be myristylated. Consistent with its myristylation and syndecan-4 association, syndesmos colocalizes with syndecan-4 in the ventral plasma membranes of cells plated on fibronectin. When overexpressed in NIH 3T3 cells, syndesmos enhances cell spreading, actin stress fiber and focal contact formation in a serum-independent manner.

scholarly journals Recent Advances and Prospects in the Research of Nascent Adhesions

2020 ◽  
Vol 11 ◽  
Author(s):  
Bernd Henning Stumpf ◽  
Andreja Ambriović-Ristov ◽  
Aleksandra Radenovic ◽  
Ana-Sunčana Smith
Keyword(s):  
Extracellular Matrix ◽  
Crucial Role ◽  
Early Stage ◽  
Focal Adhesions ◽  
Scientific Activity ◽  
Recent Advances ◽  
Transient Structures ◽  
And Function

Nascent adhesions are submicron transient structures promoting the early adhesion of cells to the extracellular matrix. Nascent adhesions typically consist of several tens of integrins, and serve as platforms for the recruitment and activation of proteins to build mature focal adhesions. They are also associated with early stage signaling and the mechanoresponse. Despite their crucial role in sampling the local extracellular matrix, very little is known about the mechanism of their formation. Consequently, there is a strong scientific activity focused on elucidating the physical and biochemical foundation of their development and function. Precisely the results of this effort will be summarized in this article.

Myotube formation is affected by adipogenic lineage cells in a cell-to-cell contact-independent manner

2014 ◽  
Vol 324 (1) ◽  
pp. 105-114 ◽  
Author(s):  
Yuki Takegahara ◽  
Keitaro Yamanouchi ◽  
Katsuyuki Nakamura ◽  
Shin-ichi Nakano ◽  
Masugi Nishihara
Keyword(s):  
Cell Contact ◽  
Independent Manner ◽  
Myotube Formation ◽  
A Cell

scholarly journals Targeting and transport: How microtubules control focal adhesion dynamics

2012 ◽  
Vol 198 (4) ◽  
pp. 481-489 ◽  
Author(s):  
Samantha Stehbens ◽  
Torsten Wittmann
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Focal Adhesion ◽  
Molecular Mechanisms ◽  
Rho Gtpase ◽  
Focal Adhesions ◽  
Matrix Metalloproteases ◽  
Force Generation ◽  
Diverse Group ◽  
Directional Cell Migration

Directional cell migration requires force generation that relies on the coordinated remodeling of interactions with the extracellular matrix (ECM), which is mediated by integrin-based focal adhesions (FAs). Normal FA turnover requires dynamic microtubules, and three members of the diverse group of microtubule plus-end-tracking proteins are principally involved in mediating microtubule interactions with FAs. Microtubules also alter the assembly state of FAs by modulating Rho GTPase signaling, and recent evidence suggests that microtubule-mediated clathrin-dependent and -independent endocytosis regulates FA dynamics. In addition, FA-associated microtubules may provide a polarized microtubule track for localized secretion of matrix metalloproteases (MMPs). Thus, different aspects of the molecular mechanisms by which microtubules control FA turnover in migrating cells are beginning to emerge.

scholarly journals AP-1B facilitates endocytosis during cell migration

2019 ◽  
Author(s):  
Margaret Johnson Kell ◽  
Su Fen Ang ◽  
Lucy Pigati ◽  
Abby Halpern ◽  
Heike Fölsch
Keyword(s):  
Plasma Membrane ◽  
Cell Migration ◽  
Epithelial Cell ◽  
Metastatic Cancer ◽  
Basolateral Membrane ◽  
Focal Adhesions ◽  
Pcr Analysis ◽  
Collective Cell Migration ◽  
Coated Pits ◽  
Cell Protrusion

ABSTRACTThe epithelial cell-specific clathrin adaptor AP-1B has a well-established role in polarized sorting of cargos to the basolateral membrane. Here we demonstrate a novel function for AP-1B during collective cell migration of epithelial sheets. We show that AP-1B colocalized with β1 integrin in focal adhesions during cell migration using confocal microscopy and total internal reflection fluorescence (TIRF) microscopy on fixed specimens. Further, AP-1B labeling in cell protrusion was distinct from labeling for the canonical endocytic adaptor complex AP-2. Using stochastic optical reconstruction microscopy (STORM) and live TIRF imaging we identified numerous AP-1B-coated structures at or close to the plasma membrane in cell protrusions. Importantly, immuno-electron microscopy (EM) showed AP-1B in clathrin-coated pits and budding vesicles at the plasma membrane during cell migration. Our data therefore established a novel function for AP-1B in endocytosis. We further show that β1 integrin was dependent on AP-1B and its co-adaptor, autosomal recessive hypercholesterolemia protein (ARH), for sorting to the basolateral membrane. Notably, we found that expression of AP-1B (and ARH) slowed epithelial-cell migration, and qRT-PCR analysis of human epithelial-derived cell lines revealed a loss of AP-1B expression in highly metastatic cancer cells indicating that AP-1B-facilitated endocytosis during cell migration might be an anti-cancer mechanism.

scholarly journals A Layered View on Focal Adhesions

Biology ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1189
Author(s):  
Karin Legerstee ◽  
Adriaan Houtsmuller
Keyword(s):  
Extracellular Matrix ◽  
Plasma Membrane ◽  
Cell Migration ◽  
Data Analysis ◽  
Protein Dynamics ◽  
Intracellular Transport ◽  
Imaging Techniques ◽  
Focal Adhesions ◽  
Potential Impact ◽  
Data Analysis Methods

The cytoskeleton provides structure to cells and supports intracellular transport. Actin fibres are crucial to both functions. Focal Adhesions (FAs) are large macromolecular multiprotein assemblies at the ends of specialised actin fibres linking these to the extracellular matrix. FAs translate forces on actin fibres into forces contributing to cell migration. This review will discuss recent insights into FA protein dynamics and their organisation within FAs, made possible by advances in fluorescence imaging techniques and data analysis methods. Over the last decade, evidence has accumulated that FAs are composed of three layers parallel to the plasma membrane. We focus on some of the most frequently investigated proteins, two from each layer, paxillin and FAK (bottom, integrin signalling layer), vinculin and talin (middle, force transduction layer) and zyxin and VASP (top, actin regulatory layer). Finally, we discuss the potential impact of this layered nature on different aspects of FA behaviour.

scholarly journals A Cell-free System to Study Regulation of Focal Adhesions and of the Connected Actin Cytoskeleton

1999 ◽  
Vol 10 (2) ◽  
pp. 373-391 ◽  
Author(s):  
Anna Cattelino ◽  
Chiara Albertinazzi ◽  
Mario Bossi ◽  
David R. Critchley ◽  
Ivan de Curtis
Keyword(s):  
Actin Cytoskeleton ◽  
Actin Polymerization ◽  
Plasma Membranes ◽  
Molecular Mechanisms ◽  
Divalent Cations ◽  
Focal Adhesions ◽  
Free System ◽  
Β1 Integrins ◽  
A Cell ◽  
And Function

Assembly and modulation of focal adhesions during dynamic adhesive processes are poorly understood. We describe here the use of ventral plasma membranes from adherent fibroblasts to explore mechanisms regulating integrin distribution and function in a system that preserves the integration of these receptors into the plasma membrane. We find that partial disruption of the cellular organization responsible for the maintenance of organized adhesive sites allows modulation of integrin distribution by divalent cations. High Ca2+ concentrations induce quasi-reversible diffusion of β1 integrins out of focal adhesions, whereas low Ca2+ concentrations induce irreversible recruitment of β1 receptors along extracellular matrix fibrils, as shown by immunofluorescence and electron microscopy. Both effects are independent from the presence of actin stress fibers in this system. Experiments with cells expressing truncated β1 receptors show that the cytoplasmic portion of β1 is required for low Ca2+-induced recruitment of the receptors to matrix fibrils. Analysis with function-modulating antibodies indicates that divalent cation-mediated receptor distribution within the membrane correlates with changes in the functional state of the receptors. Moreover, reconstitution experiments show that purified α-actinin colocalizes and redistributes with β1 receptors on ventral plasma membranes depleted of actin, implicating binding of α-actinin to the receptors. Finally, we found that recruitment of exogenous actin is specifically restricted to focal adhesions under conditions in which new actin polymerization is inhibited. Our data show that the described system can be exploited to investigate the mechanisms of integrin function in an experimental setup that permits receptor redistribution. The possibility to uncouple, under cell-free conditions, events involved in focal adhesion and actin cytoskeleton assembly should facilitate the comprehension of the underlying molecular mechanisms.

scholarly journals Extracellular matrix-specific focal adhesions in vascular smooth muscle produce mechanically active adhesion sites

2008 ◽  
Vol 295 (1) ◽  
pp. C268-C278 ◽  
Author(s):  
Zhe Sun ◽  
Luis A. Martinez-Lemus ◽  
Michael A. Hill ◽  
Gerald A. Meininger
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Mechanical Activity ◽  
Tissue Blood Flow ◽  
Cell Contact ◽  
Type I ◽  
Adhesion Sites

Integrin-mediated mechanotransduction in vascular smooth muscle cells (VSMCs) plays an important role in the physiological control of tissue blood flow and vascular resistance. To test whether force applied to specific extracellular matrix (ECM)-integrin interactions could induce myogenic-like mechanical activity at focal adhesion sites, we used atomic force microscopy (AFM) to apply controlled forces to specific ECM adhesion sites on arteriolar VSMCs. The tip of AFM probes were fused with a borosilicate bead (2∼5 μm) coated with fibronectin (FN), collagen type I (CNI), laminin (LN), or vitronectin (VN). ECM-coated beads induced clustering of α5- and β3-integrins and actin filaments at sites of bead-cell contact indicative of focal adhesion formation. Step increases of an upward ( z-axis) pulling force (800∼1,600 pN) applied to the bead-cell contact site for FN-specific focal adhesions induced a myogenic-like, force-generating response from the VSMC, resulting in a counteracting downward pull by the cell. This micromechanical event was blocked by cytochalasin D but was enhanced by jasplakinolide. Function-blocking antibodies to α5β1- and αvβ3-integrins also blocked the micromechanical cell event in a concentration-dependent manner. Similar pulling experiments with CNI, VN, or LN failed to induce myogenic-like micromechanical events. Collectively, these results demonstrate that mechanical force applied to integrin-FN adhesion sites induces an actin-dependent, myogenic-like, micromechanical event. Focal adhesions formed by different ECM proteins exhibit different mechanical characteristics, and FN appears of particular relevance in its ability to strongly attach to VSMCs and to induce myogenic-like, force-generating reactions from sites of focal adhesion in response to externally applied forces.

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