scholarly journals Focal adhesion kinase suppresses Rho activity to promote focal adhesion turnover

2000 ◽  
Vol 113 (20) ◽  
pp. 3673-3678 ◽  
Author(s):  
X.D. Ren ◽  
W.B. Kiosses ◽  
D.J. Sieg ◽  
C.A. Otey ◽  
D.D. Schlaepfer ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Adhesion ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Small Gtpase ◽  
Embryonic Lethality ◽  
Extracellular Matrices ◽  
Constitutive Activation ◽  
Focal Adhesion Turnover

Focal adhesion kinase (FAK) is activated and localized at focal adhesions upon cell adhesion to extracellular matrices. Cells lacking FAK show increased focal adhesion number and decreased cell migration, functions that are regulated by the small GTPase Rho. We now report that fibroblasts from FAK-/- mice failed to transiently inhibit Rho activity when plated on fibronectin. Re-expression of FAK restored normal Rho regulation. Turnover of focal adhesions correlated inversely with Rho activity. The presence or absence of FAK was mimicked by inhibiting or activating Rho, respectively. These data suggest that loss of FAK resulting in constitutive activation of Rho and inhibition of focal adhesion turnover can account for deficiencies in cell migration and embryonic lethality of the FAK knockout.

scholarly journals Contractility modulates cell adhesion strengthening through focal adhesion kinase and assembly of vinculin-containing focal adhesions

2010 ◽  
pp. n/a-n/a ◽  
Author(s):  
David W. Dumbauld ◽  
Heungsoo Shin ◽  
Nathan D. Gallant ◽  
Kristin E. Michael ◽  
Harish Radhakrishna ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Focal Adhesions

scholarly journals A synthetic peptide from the heparin-binding domain III (repeats III4-5) of fibronectin promotes stress-fibre and focal-adhesion formation in melanoma cells

2003 ◽  
Vol 371 (2) ◽  
pp. 565-571 ◽  
Author(s):  
José V. MOYANO ◽  
Alfredo MAQUEDA ◽  
Juan P. ALBAR ◽  
Angeles GARCIA-PARDO
Keyword(s):  
Cell Adhesion ◽  
Focal Adhesion ◽  
Adhesion Formation ◽  
Focal Adhesions ◽  
Melanoma Cells ◽  
Small Gtpase ◽  
Binding Domain ◽  
Heparin Binding ◽  
Chondroitin Sulphate Proteoglycans ◽  
Heparin Binding Domain

Cell adhesion to fibronectin results in formation of actin stress fibres and focal adhesions. In fibroblasts, this response requires two co-operative signals provided by interactions of the RGD sequence with α5β1 integrin and the heparin-binding domain II (Hep II) domain with syndecan-4. Within Hep II, this activity was mapped to repeat III13 and to the peptide FN-C/H-V(WQPPRARITGY, repeat III14). We previously described that the synthetic heparin-binding peptide/III5 (HBP/III5) (WTPPRAQITGYRLTVGLTRR, repeat III5) binds heparin and mediates cell adhesion via chondroitin sulphate proteoglycans. We have now studied whether HBP/III5 co-operates with α5β1 and drives a full cytoskeletal response in melanoma cells. SKMEL-178 cells attached and spread on the RGD-containing FNIII7–FNIII10 (FNIII7–10) fragment, but did not form stress fibres or focal adhesions. Co-immobilization of HBP/III5 with FNIII7–10 or adding soluble HBP/III5 to cells prespread on FNIII7–10, effectively induced these structures. Cell transfection with dominant-negative N19RhoA, a member of the small GTPase family, abolished the HBP/III5 effect. Both chondroitinase and heparitinase diminished focal adhesions, indicating that both types of proteoglycans bound HBP/III5 in melanoma cells. We have mapped the active sequence of HBP/III5 to YRLTVGLTRR, which is a novel sequence in fibronectin with focal-adhesion-promoting activity. The last two arginine (R) residues of this sequence are required for activity, since their replacement by alanine completely abrogated the HBP/III5 cytoskeletal effect. Moreover, this sequence is also active in the context of large fibronectin fragments. Our results establish that the Hep III region provides co-operative signals to α5β1 for the progression of the cytoskeletal response and that these include activation of RhoA.

scholarly journals Focal Adhesion Kinase Modulates Cell Adhesion Strengthening via Integrin Activation

2009 ◽  
Vol 20 (9) ◽  
pp. 2508-2519 ◽  
Author(s):  
Kristin E. Michael ◽  
David W. Dumbauld ◽  
Kellie L. Burns ◽  
Steven K. Hanks ◽  
Andrés J. García
Keyword(s):  
Cell Migration ◽  
Cell Adhesion ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Human Fibroblasts ◽  
Inducible Promoter ◽  
Time Dependent ◽  
Integrin Activation ◽  
Nonreceptor Tyrosine Kinase ◽  
Adhesive Forces

Focal adhesion kinase (FAK) is an essential nonreceptor tyrosine kinase regulating cell migration, adhesive signaling, and mechanosensing. Using FAK-null cells expressing FAK under an inducible promoter, we demonstrate that FAK regulates the time-dependent generation of adhesive forces. During the early stages of adhesion, FAK expression in FAK-null cells enhances integrin activation to promote integrin binding and, hence, the adhesion strengthening rate. Importantly, FAK expression regulated integrin activation, and talin was required for the FAK-dependent effects. A role for FAK in integrin activation was confirmed in human fibroblasts with knocked-down FAK expression. The FAK autophosphorylation Y397 site was required for the enhancements in adhesion strengthening and integrin-binding responses. This work demonstrates a novel role for FAK in integrin activation and the time-dependent generation of cell–ECM forces.

scholarly journals Src-Mediated Phosphorylation of Focal Adhesion Kinase Couples Actin and Adhesion Dynamics to Survival Signaling

2004 ◽  
Vol 24 (18) ◽  
pp. 8113-8133 ◽  
Author(s):  
M. A. Westhoff ◽  
B. Serrels ◽  
V. J. Fincham ◽  
M. C. Frame ◽  
N. O. Carragher
Keyword(s):  
Cell Migration ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Signal Transmission ◽  
Focal Adhesions ◽  
Dynamic Regulation ◽  
Anchorage Independent Growth ◽  
Biological Responses ◽  
Normal Adhesion ◽  
Survival Signaling

ABSTRACT Integrin-associated focal adhesions not only provide adhesive links between cellular actin and extracellular matrix but also are sites of signal transmission into the cell interior. Many cell responses signal through focal adhesion kinase (FAK), often by integrin-induced autophosphorylation of FAK or phosphorylation by Src family kinases. Here, we used an interfering FAK mutant (4-9F-FAK) to show that Src-dependent FAK phosphorylation is required for focal adhesion turnover and cell migration, by controlling assembly of a calpain 2/FAK/Src/p42ERK complex, calpain activation, and proteolysis of FAK. Expression of 4-9F-FAK in FAK-deficient fibroblasts also disrupts F-actin assembly associated with normal adhesion and spreading. In addition, we found that FAK's ability to regulate both assembly and disassembly of the actin and adhesion networks may be linked to regulation of the protease calpain. Surprisingly, we also found that the same interfering 4-9F-FAK mutant protein causes apoptosis of serum-deprived, transformed cells and suppresses anchorage-independent growth. These data show that Src-mediated phosphorylation of FAK acts as a pivotal regulator of both actin and adhesion dynamics and survival signaling, which, in turn, control apparently distinct processes such as cell migration and anchorage-independent growth. This also highlights that dynamic regulation of actin and adhesions (which include the integrin matrix receptors) is critical to signaling output and biological responses.

A PLCδ1-binding protein, p122RhoGAP, is localized in focal adhesions

2004 ◽  
Vol 32 (6) ◽  
pp. 1107-1109 ◽  
Author(s):  
K. Kawai ◽  
M. Yamaga ◽  
Y. Iwamae ◽  
M. Kiyota ◽  
H. Kamata ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Rat Kidney ◽  
Focal Adhesions ◽  
Small Gtpase ◽  
Cellular Distribution ◽  
Kidney Cells ◽  
Gtpase Activating Protein ◽  
Immunofluorescence Studies ◽  
Normal Rat

We have investigated the cellular distribution of p122RhoGAP, a GTPase-activating protein of Rho small GTPase and an activator of phospholipase C-δ1. Immunofluorescence studies demonstrated that endogenous p122 is localized at the tips of actin stress fibres and co-localizes with vinculin in normal rat kidney cells. In immunoprecipitation studies, p122 co-precipitated with vinculin, indicating that p122 is localized at the sites of focal adhesion. We have also shown that the N-terminal half of p122 is responsible for this localization. It is conceivable, therefore, that p122 is involved in the reorganization of the actin cytoskeleton and focal adhesions that regulate cell–substratum adhesion and cell migration.

scholarly journals Interaction of Paxillin with Poly(A)-Binding Protein 1 and Its Role in Focal Adhesion Turnover and Cell Migration

2005 ◽  
Vol 25 (9) ◽  
pp. 3763-3773 ◽  
Author(s):  
Alison J. Woods ◽  
Theodoros Kantidakis ◽  
Hisataka Sabe ◽  
David R. Critchley ◽  
Jim C. Norman
Keyword(s):  
Cell Migration ◽  
Rna Interference ◽  
Focal Adhesion ◽  
Nuclear Export ◽  
Binding Protein ◽  
Focal Adhesions ◽  
Nuclear Accumulation ◽  
Two Dimensional ◽  
Nucleocytoplasmic Shuttling ◽  
Focal Adhesion Turnover

ABSTRACT We have previously identified poly(A)-binding protein 1 (PABP1) as a ligand for paxillin and shown that the paxillin-PABP1 complex undergoes nucleocytoplasmic shuttling. By targeting the paxillin-binding subdomain sequences in PABP1, we have generated mutants of PABP1 that do not bind to cellular paxillin. Here we report that paxillin association is necessary for efficient nuclear export of PABP1 and that RNA interference of paxillin drives the nuclear accumulation of PABP1. Furthermore, ablation of paxillin-PABP1 association impeded a number of indices of cell motility including spreading on fibronectin, cell migration on two-dimensional matrices, and transmigration in Boyden chambers. These data indicate that PABP1 must associate with paxillin in order to be efficiently transported from the nucleus to the cytoplasm and that this event is necessary for cells to remodel their focal adhesions during cell migration.

scholarly journals Microtubule acetylation but not detyrosination promotes focal adhesion dynamics and cell migration

2018 ◽  
Author(s):  
Bertille Bance ◽  
Shailaja Seetharaman ◽  
Cécile Leduc ◽  
Batiste Boëda ◽  
Sandrine Etienne-Manneville
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Cell Polarization ◽  
Post Translational Modifications ◽  
Tubulin Acetylation ◽  
Focal Adhesion Turnover ◽  
Regulatory Functions ◽  
Insight Into

AbstractMicrotubules play a crucial role in mesenchymal migration by controlling cell polarity and the turnover of cell adhesive structures on the extracellular matrix. The polarized functions of microtubules imply that microtubules are locally regulated. Here, we investigated the regulation and role of two major tubulin post-translational modifications, acetylation and detyrosination, which have been associated with stable microtubules. Using primary astrocytes in a wound healing assay, we show that these tubulin modifications are independently regulated during cell polarization and differently affect cell migration. In contrast to microtubule detyrosination, αTAT1-mediated microtubule acetylation increases in the vicinity of focal adhesions and promotes cell migration. We further demonstrate that αTAT1 increases focal adhesion turnover by promoting Rab6-positive vesicle fusion at focal adhesions. Our results highlight the specificity of microtubule post-translational modifications and bring new insight into the regulatory functions of tubulin acetylation.

scholarly journals Structural Insight into the Mechanisms of Targeting and Signaling of Focal Adhesion Kinase

2002 ◽  
Vol 22 (8) ◽  
pp. 2751-2760 ◽  
Author(s):  
Gaohua Liu ◽  
Cristina D. Guibao ◽  
Jie Zheng
Keyword(s):  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Solution Structure ◽  
Hydrophobic Interactions ◽  
Focal Adhesions ◽  
Nonreceptor Tyrosine Kinase ◽  
Binding Partner ◽  
Focal Adhesion Turnover ◽  
Structural Insight ◽  
Insight Into

ABSTRACT Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase whose focal adhesion targeting (FAT) domain interacts with other focal adhesion molecules in integrin-mediated signaling. Localization of activated FAK to focal adhesions is indispensable for its function. Here we describe a solution structure of the FAT domain bound to a peptide derived from paxillin, a FAK-binding partner. The FAT domain is composed of four helices that form a “right-turn” elongated bundle; the globular fold is mainly maintained by hydrophobic interactions. The bound peptide further stabilizes the structure. Certain signaling events such as phosphorylation and molecule interplay may induce opening of the helix bundle. Such conformational change is proposed to precede departure of FAK from focal adhesions, which starts focal adhesion turnover.

scholarly journals Focal adhesion kinase confers pro-migratory and anti-apoptotic properties and is a potential therapeutic target in Ewing sarcoma

2019 ◽  
Author(s):  
Konrad Steinestel ◽  
Esther-Pia Jansen ◽  
Marcel Trautmann ◽  
Uta Dirksen ◽  
Jan Rehkämper ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Ewing Sarcoma ◽  
Chorioallantoic Membrane ◽  
Focal Adhesions ◽  
Childhood And Adolescence ◽  
Tissue Samples

ABSTRACTOncogenesis of Ewing sarcoma (EwS), the second most common malignant bone tumor of childhood and adolescence, is dependent on the expression of chimeric EWSR1-ETS fusion oncogenes, most often EWSR1-FLI1 (E/F).E/F expression leads to dysregulation of focal adhesions (FAs) enhancing the migratory capacity of EwS cells. Here we show that, in EwS cell lines and tissue samples, focal adhesion kinase (FAK) is expressed and phosphorylated at Y397 in an E/F-dependent way involving Ezrin. Employing different EwS cell as in vitro models, we found that key malignant properties of E/F are mediated via substrate-independent autophosphorylation of FAK on Y397. This phosphorylation results in enhanced FA formation, Rho-dependent cell migration, and impaired caspase-3-mediated apoptosis in vitro. Conversely, treatment with the FAK inhibitor Y15 enhanced caspase-mediated apoptosis and EwS cell migration, independent from the respective EWSR1-ETS fusion type, mimicking an anoikis-like phenotype. Our findings were confirmed in vivo using an avian chorioallantoic membrane (CAM) model. Our results provide a first rationale for the therapeutic use of FAK inhibitors to impair metastatic dissemination of EwS.

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