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

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.

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Faculty Opinions recommendation of Structural insight into the mechanisms of targeting and signaling of focal adhesion kinase.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1005457.64755 ◽

2002 ◽

Author(s):

Christopher Turner

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Structural Insight ◽

Insight Into

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

10.1101/425421 ◽

2018 ◽

Cited By ~ 1

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.

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Faculty Opinions recommendation of Structural insight into the mechanisms of targeting and signaling of focal adhesion kinase.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1005457.66218 ◽

2002 ◽

Author(s):

Michael Schaller

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Structural Insight ◽

Insight Into

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Focal adhesion kinase suppresses Rho activity to promote focal adhesion turnover

Journal of Cell Science ◽

10.1242/jcs.113.20.3673 ◽

2000 ◽

Vol 113 (20) ◽

pp. 3673-3678 ◽

Cited By ~ 27

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.

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Insight into the HIV-1 Vif SOCS-box–ElonginBC interaction

Open Biology ◽

10.1098/rsob.130100 ◽

2013 ◽

Vol 3 (11) ◽

pp. 130100 ◽

Cited By ~ 7

Author(s):

Zhisheng Lu ◽

Julien R. C. Bergeron ◽

R. Andrew Atkinson ◽

Torsten Schaller ◽

Dennis A. Veselkov ◽

...

Keyword(s):

Solution Structure ◽

Hydrophobic Interactions ◽

Dynamic Information ◽

Ubiquitin Ligase Complex ◽

Biophysical Studies ◽

Viral Infectivity Factor ◽

Β Sheet ◽

Socs Box ◽

Insight Into ◽

Hiv 1

The HIV-1 viral infectivity factor (Vif) neutralizes cell-encoded antiviral APOBEC3 proteins by recruiting a cellular ElonginB (EloB)/ElonginC (EloC)/Cullin5-containing ubiquitin ligase complex, resulting in APOBEC3 ubiquitination and proteolysis. The suppressors-of-cytokine-signalling-like domain (SOCS-box) of HIV-1 Vif is essential for E3 ligase engagement, and contains a BC box as well as an unusual proline-rich motif. Here, we report the NMR solution structure of the Vif SOCS–ElonginBC (EloBC) complex. In contrast to SOCS-boxes described in other proteins, the HIV-1 Vif SOCS-box contains only one α-helical domain followed by a β-sheet fold. The SOCS-box of Vif binds primarily to EloC by hydrophobic interactions. The functionally essential proline-rich motif mediates a direct but weak interaction with residues 101–104 of EloB, inducing a conformational change from an unstructured state to a structured state. The structure of the complex and biophysical studies provide detailed insight into the function of Vif's proline-rich motif and reveal novel dynamic information on the Vif–EloBC interaction.

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Molecular basis for Ras suppressor-1 recruitment to focal adhesions and stabilization of consensus adhesome complex

10.1101/2021.02.19.432017 ◽

2021 ◽

Author(s):

Koichi Fukuda ◽

Fan Lu ◽

Jun Qin

Keyword(s):

Cell Adhesion ◽

Focal Adhesion ◽

Specific Interaction ◽

Tumor Development ◽

Focal Adhesions ◽

Adaptor Protein ◽

Biological Data ◽

Structural Basis ◽

Lim Domain ◽

Insight Into

AbstractRas suppressor-1 (Rsu-1) is a leucine-rich repeat (LRR)-containing protein that is crucial for regulating fundamental cell adhesion processes and tumor development. Rsu-1 interacts with a zinc-finger type multi LIM domain-containing adaptor protein PINCH-1 involved in the integrin-mediated consensus adhesome but not with highly homologous isoform PINCH-2. However, the structural basis for such specific interaction and regulatory mechanism remains unclear. Here, we determined the crystal structures of Rsu-1 and its complex with the PINCH-1 LIM4-5 domains. Rsu-1 displays an arc-shaped solenoid architecture with eight LRRs shielded by the N- and C-terminal capping modules. We show that a large conserved concave surface of the Rsu-1 LRR domain recognizes the PINCH-1 LIM5 domain, and that the C-terminal non-LIM region of PINCH-2 but not PINCH-1 sterically disfavors the Rsu-1 binding. We further show that Rsu-1 can be assembled, via PINCH-1-binding, into a tight hetero-pentamer complex comprising Rsu-1, PINCH-1, ILK, Parvin, and Kindlin-2 that constitute a major consensus integrin adhesome crucial for focal adhesion assembly. Consistently, our mutagenesis and cell biological data consolidate the significance of the Rsu-1/PINCH-1 interaction in focal adhesion assembly and cell spreading. Our results provide a crucial molecular insight into Rsu-1-mediated cell adhesion with implication on how it may regulate tumorigenic growth.

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Cardiac fibroblasts require focal adhesion kinase for normal proliferation and migration

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00444.2008 ◽

2009 ◽

Vol 296 (3) ◽

pp. H627-H638 ◽

Cited By ~ 17

Author(s):

Ana Maria Manso ◽

Seok-Min Kang ◽

Sergey V. Plotnikov ◽

Ingo Thievessen ◽

Jaewon Oh ◽

...

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Adult Mouse ◽

Cardiac Fibroblasts ◽

Focal Adhesions ◽

Protein Levels ◽

Tyrosine Kinase 2 ◽

A Cell ◽

And Migration ◽

Proliferation And Migration

Migration and proliferation of cardiac fibroblasts (CFs) play an important role in the myocardial remodeling process. While many factors have been identified that regulate CF growth and migration, less is known about the signaling mechanisms involved in these processes. Here, we utilized Cre-LoxP technology to obtain focal adhesion kinase (FAK)-deficient adult mouse CFs and studied how FAK functioned in modulating cell adhesion, proliferation, and migration of these cells. Treatment of FAKflox/flox CFs with Ad/Cre virus caused over 70% reduction of FAK protein levels within a cell population. FAK-deficient CFs showed no changes in focal adhesions, cell morphology, or protein expression levels of vinculin, talin, or paxillin; proline-rich tyrosine kinase 2 (Pyk2) expression and activity were increased. Knockdown of FAK protein in CFs increased PDGF-BB-induced proliferation, while it reduced PDGF-BB-induced migration. Adhesion to fibronectin was not altered. To distinguish between the function of FAK and Pyk2, FAK function was inhibited via adenoviral-mediated overexpression of the natural FAK inhibitor FAK-related nonkinase (FRNK). Ad/FRNK had no effect on Pyk2 expression, inhibited the PDGF-BB-induced migration, but did not change the PDGF-BB-induced proliferation. FAK deficiency had only modest effects on increasing PDGF-BB activation of p38 and JNK MAPKs, with no alteration in the ERK response vs. control cells. These results demonstrate that FAK is required for the PDGF-BB-induced migratory response of adult mouse CFs and suggest that FAK could play an essential role in the wound-healing response that occurs in numerous cardiac pathologies.

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