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 IFT20 Mediates the Transport of Cell Migration Regulators From the Trans-Golgi Network to the Plasma Membrane in Breast Cancer Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Huihui Yang ◽  
Fan Zhang ◽  
Huan Long ◽  
Yiwen Lin ◽  
Jiahui Liao ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Plasma Membrane ◽  
Cell Migration ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Intracellular Transport ◽  
Primary Cilia ◽  
Intraflagellar Transport ◽  
Trans Golgi Network ◽  
Golgi Network

IFT20 is a subunit of the intraflagellar transport (IFT) system essential for the formation and function of cilia. Besides predominant research in the cilia field, some IFT subunits perform extraciliary roles in non-ciliated cancer cells. However, the specific roles of IFT subunits in tumorigenesis remain unknown. Here, we found that knockout of IFT20 in mouse breast cancer cells lacking primary cilia promoted epithelial mesenchymal transitions (EMTs), active lamellipodia formation, and cell migration. IFT20 localized at the trans-Golgi and trans-Golgi network (TGN), and displayed vesicular co-distributions with Rab8a, the marker of TGN-to-plasma membrane vesicular trafficking. Proximity-dependent biotin identification (BioID) and colocalization analyzes showed that Numb and Ctnnal1, whose depletion promoted cell migration, co-localized with IFT20 at the trans-Golgi/TGN or intracellular transport vesicles. Furthermore, Strep-Tactin pulldown assays revealed an interaction between IFT20 and Ctnnal1 or Numb. Loss of IFT20 lowered the expression of actin-associated Tagln2, whose knockdown promoted cell migration. Thus, the extraciliary function of ITF20 in breast cancer cell was associated with the negative regulation of migration.

scholarly journals Calcium-stimulated disassembly of focal adhesions mediated by an ORP3/IQSec1 complex

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Ryan S D'Souza ◽  
Jun Y Lim ◽  
Alper Turgut ◽  
Kelly Servage ◽  
Junmei Zhang ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Migration ◽  
Calcium Channels ◽  
Lipid Transfer Protein ◽  
Calcium Influx ◽  
Focal Adhesions ◽  
Lipid Transfer ◽  
Transfer Protein ◽  
Contact Sites ◽  
Lipid Exchange

Coordinated assembly and disassembly of integrin-mediated focal adhesions (FAs) is essential for cell migration. Many studies have shown that FA disassembly requires Ca2+ influx, however our understanding of this process remains incomplete. Here, we show that Ca2+ influx via STIM1/Orai1 calcium channels, which cluster near FAs, leads to activation of the GTPase Arf5 via the Ca2+-activated GEF IQSec1, and that both IQSec1 and Arf5 activation are essential for adhesion disassembly. We further show that IQSec1 forms a complex with the lipid transfer protein ORP3, and that Ca2+ influx triggers PKC-dependent translocation of this complex to ER/plasma membrane (PM) contact sites adjacent to FAs. In addition to allosterically activating IQSec1, ORP3 also extracts PI4P from the PM, in exchange for phosphatidylcholine. ORP3-mediated lipid exchange is also important for FA turnover. Together, these findings identify a new pathway that links calcium influx to FA turnover during cell migration.

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 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 Zyxin Is Involved in Fibroblast Rigidity Sensing and Durotaxis

2021 ◽  
Vol 9 ◽  
Author(s):  
Ai Kia Yip ◽  
Songjing Zhang ◽  
Lor Huai Chong ◽  
Elsie Cheruba ◽  
Jessie Yong Xing Woon ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Focal Adhesions ◽  
Wild Type ◽  
Actin Organization ◽  
Rigidity Sensing ◽  
Substrate Rigidity ◽  
Cell Traction ◽  
Traction Stress

Focal adhesions (FAs) are specialized structures that enable cells to sense their extracellular matrix rigidity and transmit these signals to the interior of the cells, bringing about actin cytoskeleton reorganization, FA maturation, and cell migration. It is known that cells migrate towards regions of higher substrate rigidity, a phenomenon known as durotaxis. However, the underlying molecular mechanism of durotaxis and how different proteins in the FA are involved remain unclear. Zyxin is a component of the FA that has been implicated in connecting the actin cytoskeleton to the FA. We have found that knocking down zyxin impaired NIH3T3 fibroblast’s ability to sense and respond to changes in extracellular matrix in terms of their FA sizes, cell traction stress magnitudes and F-actin organization. Cell migration speed of zyxin knockdown fibroblasts was also independent of the underlying substrate rigidity, unlike wild type fibroblasts which migrated fastest at an intermediate substrate rigidity of 14 kPa. Wild type fibroblasts exhibited durotaxis by migrating toward regions of increasing substrate rigidity on polyacrylamide gels with substrate rigidity gradient, while zyxin knockdown fibroblasts did not exhibit durotaxis. Therefore, we propose zyxin as an essential protein that is required for rigidity sensing and durotaxis through modulating FA sizes, cell traction stress and F-actin organization.

scholarly journals Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration

2018 ◽  
Vol 2 (4) ◽  
pp. 046107 ◽  
Author(s):  
William Y. Wang ◽  
Alexander T. Pearson ◽  
Matthew L. Kutys ◽  
Colin K. Choi ◽  
Michele A. Wozniak ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Focal Adhesions

scholarly journals Turnover of Focal Adhesions and Cancer Cell Migration

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Makoto Nagano ◽  
Daisuke Hoshino ◽  
Naohiko Koshikawa ◽  
Toshifumi Akizawa ◽  
Motoharu Seiki
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Cancer Cell ◽  
Focal Adhesions ◽  
Cancer Cell Migration ◽  
Disassembly Process ◽  
Molecular Events ◽  
Interaction Partners ◽  
Cells Cultured

Cells are usually surrounded by the extracellular matrix (ECM), and adhesion of the cells to the ECM is a key step in their migration through tissues. Integrins are important receptors for the ECM and form structures called focal adhesions (FAs). Formation and disassembly of FAs are regulated dynamically during cell migration. Adhesion to the ECM has been studied mainly using cells cultured on an ECM-coated substratum, where the rate of cell migration is determined by the turnover of FAs. However, the molecular events underlying the disassembly of FAs are less well understood. We have recently identified both a new regulator of this disassembly process and its interaction partners. Here, we summarize our understanding of FA disassembly by focusing on the proteins implicated in this process.

scholarly journals PtdIns(4,5)P2 and PtdIns(3,4,5)P3 dynamics during focal adhesions assembly and disassembly in a cancer cell line

2020 ◽  
Vol 44 (6) ◽  
pp. 381-392
Author(s):  
Dhurgham ALFAHAD ◽  
Salem ALHARETHI ◽  
Bandar ALHARBI ◽  
Khatab MAWLOOD ◽  
Philip DASH
Keyword(s):  
Extracellular Matrix ◽  
Plasma Membrane ◽  
Cancer Cell ◽  
Membrane Lipids ◽  
Focal Adhesions ◽  
Cancer Cell Line ◽  
Cancer Cell Migration ◽  
Fluorescent Biosensors ◽  
Intracellular Signals ◽  
Local Levels

Focal adhesions (FAs) are large assemblies of proteins that mediate intracellular signals between the cytoskeleton and the extracellular matrix (ECM). The turnover of FA proteins plays a critical regulatory role in cancer cell migration. Plasma membrane lipids locally generated or broken down by different inositide kinases and phosphatase enzymes to activate and recruit proteins to specific regions in the plasma membrane. Presently, little attention has been given to the use of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and Phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) fluorescent biosensors in order to determine the spatiotemporal organisation of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 within and around or during assembly and disassembly of FAs. In this study, specific biosensors were used to detect PtdIns(4,5)P2, PtdIns(3,4,5)P3, and FAs proteins conjugated to RFP/GFP in order to monitor changes of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 levels within FAs. We demonstrated that the localisation of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 were moderately correlated with that of FA proteins. Furthermore, we demonstrate that local levels of PtdIns(4,5)P2 increased within FA assembly and declined within FA disassembly. However, PtdIns(3,4,5)P3 levels remained constant within FAs assembly and disassembly. In conclusion, this study shows that PtdIns(4,5)P2 and PtdIns(3,4,5)P3 localised in FAs may be regulated differently during FA assembly and disassembly.

scholarly journals Mitochondria Tether to Focal Adhesions During Cell Migration and Regulate Their Size.

2021 ◽  
Author(s):  
Redaet Daniel ◽  
Patricia Bilodeau ◽  
Abebech Mengeta ◽  
Kimmy Yang ◽  
Jonathan M. Lee
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Cell Motility ◽  
Focal Adhesions ◽  
Leading Edge ◽  
Integrin Receptors ◽  
Substrate Interaction ◽  
Functional Part ◽  
Atp Generation ◽  
Contractile Forces

Abstract Focal Adhesions (FA) couple the actin cytoskeleton to the extracellular matrix through transmembrane integrin receptors. FA assembly and disassembly regulate cell migration by controlling substrate interaction and the generation of intracellular contractile forces. Here we show that FA interact with mitochondria. Mitochondria are highly dynamic organelles that are now emerging as regulators of mammalian cell motility. We find that mitochondria infiltrate the leading edge of NIH3T3 fibroblasts during migration and tether to FA there. Importantly, we find that FA interacting with mitochondria are larger than those lacking mitochondrial interaction. In addition, inhibition of mitochondrial ATP generation reduces FA size and artificial tethering of FA to mitochondria concomitantly increases their size. Taken together this suggests that mitochondrial interaction with FA is a functional part of cell migration and adhesion.

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