Alternative Splicing Controls the Mechanisms of FAK Autophosphorylation

Toutant, Madeleine; Costa, Alícia; Studler, Jeanne-Marie; Kadaré, Gress; Carnaud, Michèle; Girault, Jean‐Antoine

doi:10.1128/mcb.22.22.7731-7743.2002

Cited by 130 publications

(150 citation statements)

References 59 publications

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“…It is unclear exactly how clustering of integrins leads to activation of FAK. Perhaps the clustering of integrins at sites of cell adhesion, with the subsequent localization of FAK molecules in close proximity, may lead to intermolecular tyrosine phosphorylation (Polte and Hanks, 1997;Toutant et al, 2002). Alternatively, conformational changes in FAK upon integrin binding may expose phosphorylation sites or sequestration from tyrosine phosphatases (Hanks et al, 2003).…”

Section: Src and Integrin Signalingmentioning

confidence: 99%

The interplay between Src and integrins in normal and tumor biology

2004

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Src family nonreceptor protein tyrosine kinases transduce signals that control normal cellular processes such as cell proliferation, adhesion and motility. Normally, cellular Src is held in an inactive state, but in several cancer types, abnormal events lead to elevated kinase activity of the protein and cause pleiotropic cellular responses inducing transformation and metastasis. A prerequisite of the ability of a cancer cell to undergo metastasis into distant tissues is to penetrate surrounding extracellular matrices. These processes are facilitated by the integrin family of cell adhesion molecules. As is the case with Src, altered integrin activity or substrate affinity can contribute to the neoplastic phenotype. Therefore, understanding the interplay between Src and integrin function has been of intense interest over the past few years. This review focuses on the role of Src and integrin signaling in normal cells and how this is deregulated in human cancer. We will identify the key players in the integrin-mediated signaling pathways involved in cell motility and apoptosis, such as FAK, paxillin and p130 CAS , and discuss how Src signaling affects the formation of focal adhesions and the extracellular matrix.

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Section: Src and Integrin Signalingmentioning

confidence: 99%

The interplay between Src and integrins in normal and tumor biology

2004

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“…Proteins were partly renatured by incubation in buffer (Hepes, pH 7.4, MgCl 2 10 mM, NaCl 50 mM, EDTA 1 mM, dithiothreitol 1 mM, glycerol 10%) for 24 h. After saturation of nonspecific sites with 5% nonfat dry milk, the membrane was incubated overnight with purified recombinant GST-N-FAK (75 ng/cm 2 of membrane), washed extensively, and processed for immunoblotting using anti-FAK antibody A-17. The other procedures were as described (23,30).…”

Section: Maucuer (Inserm 440 Paris)mentioning

confidence: 99%

“…Cells were lysed 48 h after transfection. Comparisons of attached and suspended cells (23) and cell partitioning (29) were as described.…”

Section: Maucuer (Inserm 440 Paris)mentioning

confidence: 99%

“…Cell Culture, Transfection, and Fractionation-COS-7 and NIH-3T3 cells were cultured and transfected with 8 g of DNA per 100-mmdiameter culture dish as described (23). For triple transfections, the amounts of transfected plasmids were: 2 g of pBK-FAK (23) or pBK-CMV3-PYK2, 2 g of pSG5-SUMO-1 (a generous gift from J. Seeler, Pasteur Institute), and the indicated amounts of pcDNA3-HA-PIAS1.…”

Section: Maucuer (Inserm 440 Paris)mentioning

confidence: 99%

“…This domain has the capacity to interact with ezrin (19) and peptides mimicking intracellular regions of ␤-integrins (20). Deletion of this domain, or part of it, increases tyrosine phosphorylation and autophosphorylation of FAK, suggesting that it regulates FAK activity (21)(22)(23).…”

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PIAS1-mediated Sumoylation of Focal Adhesion Kinase Activates Its Autophosphorylationn

Kadaré

Toutant

Formstecher

et al. 2003

Journal of Biological Chemistry

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Focal adhesion kinase (FAK) is a protein tyrosine kinase enriched in focal adhesions, which plays a critical role in integrin-dependent cell motility and survival. The crucial step in its activation is autophosphorylation on Tyr-397, which promotes the recruitment of several enzymes including Src family kinases and the activation of multiple signaling pathways. We found in a yeast two-hybrid screen that the N-terminal domain of FAK interacted with protein inhibitor of activated STAT1 (PIAS1). This interaction was confirmed and shown to be direct using in vitro assays. PIAS1 was co-immunoprecipitated with FAK from transfected cells and brain extracts. PIAS1 has recently been recognized as a small ubiquitin-like modifier (SUMO) ligase. In the presence of PIAS1 and SUMO-1, FAK was sumoylated in intact cells, whereas PYK2, a closely related enzyme, was not. Sumoylation occurred on Lys-152, a residue conserved in FAK during evolution. Sumoylated FAK, like PIAS1, was recovered predominantly from the nuclear fraction. Sumoylation did not require the catalytic activity or autophosphorylation of FAK. In contrast, sumoylation increased dramatically the ability of FAK to autophosphorylate in intact cells and in immune precipitate kinase assays. Endogenous FAK was sumoylated in the presence of PIAS1 and SUMO-1 independently of cell adhesion, and autophosphorylation of sumoylated FAK was persistently increased in suspended cells. These observations show that sumoylation controls the activity of a protein kinase and suggest that FAK may play a novel role in signaling between the plasma membrane and the nucleus.Focal adhesion kinase (FAK) 1 is a non-receptor cytoplasmic tyrosine kinase of 125 kDa (1, 2) implicated in integrin-mediated signal transduction (reviewed in Refs. 3-5). It is also activated by a variety of extracellular stimuli including G protein-coupled receptors and growth factor receptors (see Refs. 5 and 6). In adherent mammalian cells in culture FAK is located in focal adhesions (1, 2) and appears important for the regulation of their turnover (7). FAK is critical for adhesion-dependent cell survival (8) and integrin-mediated motility (9). Its physiological importance is demonstrated by the embryonic mortality of FAK knockout mice (7), whereas recent work underlines its role in tumor invasiveness (10). Autophosphorylation at Tyr-397 is a crucial event for FAK biological function, because it creates a high affinity binding site for proteins harboring Src homology 2 (SH2) domains, such as Src and Fyn (11), Grb7 (12), and phosphatidylinositide 3Ј-OH-kinase, which activates the anti-apoptotic Akt pathway (13,14). Following their binding to phospho-Tyr-397, Src or Fyn phosphorylate FAK at other tyrosine residues (15, 16) as well as associated proteins, thereby leading to the activation of several signaling pathways (see 5).In contrast to the recent progress in elucidating the signaling pathways downstream from FAK, relatively little is known about the molecular mechanisms regulating FAK activity. The tyrosine kinase ...

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Integrin β1 transduces the signal for LY6D‐induced macropinocytosis and mediates senescence‐inducing stress‐evoked vacuole formation via FAK

et al. 2022

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Cellular senescence is a highly stable cell-cycle arrest induced by DNA damage and various cellular stresses. Recently, we have revealed that lymphocyte antigen 6 complex, locus D (LY6D) is responsible for senescence-inducing stress-evoked vacuole formation through induction of Src family kinase (SFK)-mediated macropinocytosis. However, the signaling molecule(s) transducing the macropinocytosis signal from extracellular LY6D to the cytoplasmic SFK are unknown. In this study, we identified integrin b1, a transmembrane signaling protein, as an interactor of LY6D by proteomic analysis and co-immunoprecipitation assays. Inhibition of integrin b1 impaired LY6D-induced macropinocytosis, and integrin b1 activated SFK through focal adhesion kinase to mediate macropinocytosis. These results indicate that integrin b1 is a crucial mediator of the LY6D-induced vacuole formation in senescent cells.

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Alternative Splicing Controls the Mechanisms of FAK Autophosphorylation

Cited by 130 publications

References 59 publications

The interplay between Src and integrins in normal and tumor biology

The interplay between Src and integrins in normal and tumor biology

PIAS1-mediated Sumoylation of Focal Adhesion Kinase Activates Its Autophosphorylationn

Integrin β1 transduces the signal for LY6D‐induced macropinocytosis and mediates senescence‐inducing stress‐evoked vacuole formation via FAK

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