scholarly journals Paxillin null embryonic stem cells are impaired in cell spreading and tyrosine phosphorylation of focal adhesion kinase

Oncogene ◽  
2002 ◽  
Vol 21 (1) ◽  
pp. 96-107 ◽  
Author(s):  
Ramon Wade ◽  
Joanna Bohl ◽  
Scott Vande Pol
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Embryonic Stem ◽  
Cell Spreading

scholarly journals Minimal features of paxillin that are required for the tyrosine phosphorylation of focal adhesion kinase

2005 ◽  
Vol 393 (2) ◽  
pp. 565-573 ◽  
Author(s):  
Ramon Wade ◽  
Scott Vande Pol
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Embryonic Stem ◽  
Adaptor Protein ◽  
Structural Features ◽  
Lim Domains

Tyrosine phosphorylation of FAK (focal adhesion kinase) regulates signalling that results from the interaction of integrins with extracellular matrix and growth factor receptors. A critical step in this process is the phosphorylation of Tyr397 of FAK, which creates a binding site for Src family kinases, PI3K (phosphoinositide 3-kinase) and Shc (Src homology and collagen homology). An intact Tyr397 site is required for FAK-mediated regulation of cell migration, survival signals and full responsiveness to soluble growth factors. We showed previously that the adaptor protein paxillin is required for the overall tyrosine phosphorylation of FAK in embryonic stem cells [Wade, Bohl and Vande Pol (2002) Oncogene 21, 96–107]. In the present paper, we identify the minimal structural features of paxillin that are required to support overall FAK tyrosine phosphorylation and Tyr397 phosphorylation. Paxillin contains N-terminal leucine-rich LD motifs that bind directly to FAK and four LIM (Lin-11, Isl-1 and Mec-3) domains in the C-terminus. We show that paxillin LIM domains 1, 2 and 3 are each required for FAK tyrosine phosphorylation, while LIM4 is dispensable. In addition to paxillin LIM domains 1, 2 and 3, a single LD motif on paxillin is required to support FAK tyrosine phosphorylation in embryonic stem cells. Both sequence and spatial requirements exist for LD motifs to support FAK tyrosine phosphorylation. Interestingly, synthetic LD motifs that fail to bind FAK in vitro are able to fully support FAK tyrosine phosphorylation, indicating that minimal interactions of LD motifs with FAK suffice. Our results demonstrate at least four distinct structural domains of paxillin support at least three distinct functions that are each required for FAK tyrosine phosphorylation.

scholarly journals Focal Adhesion Kinase Signaling Regulates Cardiogenesis of Embryonic Stem Cells

2005 ◽  
Vol 280 (47) ◽  
pp. 39534-39544 ◽  
Author(s):  
Daihiko Hakuno ◽  
Tomosaburo Takahashi ◽  
Jan Lammerding ◽  
Richard T. Lee
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Embryonic Stem ◽  
Kinase Signaling ◽  
Focal Adhesion Kinase Signaling

scholarly journals Integrin-Associated Focal Adhesion Kinase Protects Human Embryonic Stem Cells from Apoptosis, Detachment, and Differentiation

2016 ◽  
Vol 7 (2) ◽  
pp. 167-176 ◽  
Author(s):  
Loriana Vitillo ◽  
Melissa Baxter ◽  
Banu Iskender ◽  
Paul Whiting ◽  
Susan J. Kimber
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

scholarly journals Identification of integrin-stimulated sites of serine phosphorylation in FRNK, the separately expressed C-terminal domain of focal adhesion kinase: a potential role for protein kinase A

1997 ◽  
Vol 324 (1) ◽  
pp. 141-149 ◽  
Author(s):  
Alan RICHARDSON ◽  
John D. SHANNON ◽  
Reid B. ADAMS ◽  
Michael D. SCHALLER ◽  
J. Thomas PARSONS
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Serine Phosphorylation ◽  
Early Event ◽  
Kinase A

Focal adhesion kinase (pp125FAK) is a protein tyrosine kinase that is localized to focal adhesions in many cell types and which undergoes tyrosine phosphorylation after integrin binding to extracellular matrix. In some cells the C-terminal non-catalytic domain of pp125FAK is expressed as a separate protein referred to as FRNK (FAK-related, non-kinase). We have previously shown that overexpression of FRNK inhibits tyrosine phosphorylation of pp125FAK and its substrates as well as inhibiting cell spreading on fibronectin. In this report we identify Ser148 and Ser151 as residues in FRNK that are phosphorylated after tyrosine phosphorylation of pp125FAK and in response to integrin binding to fibronectin. Tyrosine phosphorylation of pp125FAK appears to be an early event after integrin occupancy, and serine phosphorylation of FRNK occurs significantly later. Treatment of fibroblasts with a series of protein kinase A inhibitors delayed serine phosphorylation of FRNK as well as cell spreading on fibronectin and tyrosine phosphorylation of pp125FAK. However, these PKA inhibitors are unlikely to delay cell spreading simply by preventing serine phosphorylation of FRNK, as overexpression of FRNK containing mutations of Ser148 and Ser151 either singly or jointly to either alanine or glutamate residues did not significantly alter the ability of FRNK to act as an inhibitor of pp125FAK.

Low Focal Adhesion Signaling Promotes Ground State Pluripotency of Mouse Embryonic Stem Cells

2017 ◽  
Vol 16 (10) ◽  
pp. 3585-3595 ◽  
Author(s):  
Sara Taleahmad ◽  
Mehdi Mirzaei ◽  
Azam Samadian ◽  
Seyedeh-Nafiseh Hassani ◽  
Paul A. Haynes ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Focal Adhesion ◽  
Ground State ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells

scholarly journals The Cytoplasmic Tyrosines of Integrin Subunit β1 Are Involved in Focal Adhesion Kinase Activation

2000 ◽  
Vol 20 (15) ◽  
pp. 5758-5765 ◽  
Author(s):  
Krister Wennerberg ◽  
Annika Armulik ◽  
Takao Sakai ◽  
Marjam Karlsson ◽  
Reinhard Fässler ◽  
...  
Keyword(s):  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Phosphorylation Site ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Integrin Subunit ◽  
Wild Type ◽  
Directed Cell Migration ◽  
Dependent Cell

ABSTRACT We have previously shown that mutation of the two tyrosines in the cytoplasmic domain of integrin subunit β1 (Y783 and Y795) to phenylalanines markedly reduces the capability of β1A integrins to mediate directed cell migration. In this study, β1-dependent cell spreading was found to be delayed in GD25 cells expressing β1AY783/795F compared to that in wild-type GD25-β1A. Focal adhesion kinase (FAK) tyrosine phosphorylation and activation were severely impaired in response to β1-dependent adhesion in GD25-β1AY783/795F cells compared to that in wild-type GD25-β1A or mutants in which only a single tyrosine was altered (β1AY783F or β1AY795F). Phosphorylation site-specific antibodies selective for FAK phosphotyrosine 397 indicated that the defect in FAK phosphorylation via β1AY783/795F lies at the level of the initial autophosphorylation step. Indeed, β1A-dependent tyrosine phosphorylation of tensin and paxillin was lost in the β1AY783/795F cells, consistent with the impairment in FAK activation. In contrast, p130CAS overall tyrosine phosphorylation was unaffected by the β1 mutations. Despite the defect in β1-mediated FAK activation, FAK was still localized to focal adhesions. Taken together, the phenotype of the GD25-β1AY783/795F cells resembles, but is distinct from, the phenotype observed in FAK-null cells. These observations argue that tyrosines 783 and 795 within the cytoplasmic tail of integrin subunit β1A are critical mediators of FAK activation and cell spreading in GD25 cells.

scholarly journals Tyrosine Phosphorylation Profiling in FGF-2 Stimulated Human Embryonic Stem Cells

PLoS ONE ◽  
2011 ◽  
Vol 6 (3) ◽  
pp. e17538 ◽  
Author(s):  
Vanessa M. Y. Ding ◽  
Paul J. Boersema ◽  
Leong Yan Foong ◽  
Christian Preisinger ◽  
Geoffrey Koh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Tyrosine Phosphorylation ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

scholarly journals The self-renewal of mouse embryonic stem cells is regulated by cell–substratum adhesion and cell spreading

2013 ◽  
Vol 45 (11) ◽  
pp. 2698-2705 ◽  
Author(s):  
Patricia Murray ◽  
Marina Prewitz ◽  
Isabel Hopp ◽  
Nicola Wells ◽  
Haifei Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cell Spreading ◽  
Mouse Embryonic Stem Cells ◽  
The Self ◽  
Self Renewal

Focal adhesion kinase is redistributed to focal complexes and mediates cell spreading in macrophages in response to M-CSF

2005 ◽  
Vol 386 (9) ◽  
Author(s):  
Elisabetta Rovida ◽  
Benedetta Lugli ◽  
Valentina Barbetti ◽  
Serena Giuntoli ◽  
Massimo Olivotto ◽  
...  
Keyword(s):  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Tyrosine Kinases ◽  
Active Form ◽  
Cell Spreading ◽  
Dominant Negative ◽  
Mononuclear Phagocytes ◽  
Relevant Role ◽  
Macrophage Colony

AbstractThe macrophage colony-stimulating factor (M-CSF, CSF-1) regulates survival, proliferation and differentiation of mononuclear phagocytes, as well as macrophage motility and morphology. The latter features are usually regulated by ECM-mediated activation of integrins and subsequent tyrosine phosphorylation of cellular proteins, including focal adhesion kinase (FAK). FAK is phosphorylated by downstream receptor tyrosine kinases as well. We addressed the question whether M-CSF regulates FAK tyrosine phosphorylation in macrophages, and found that M-CSF induces FAK phosphorylation at all known tyrosine residues. This phosphorylation was dependent on Src. Extracellularly-regulated kinase (ERK), Jun N-terminal kinase (JNK) and phosphatidylinositol-3-kinase (PI3K) were found to be negatively involved in M-CSF-induced FAK phosphorylation, as their inhibition resulted in FAK hyper-phosphorylation. Following M-CSF treatment, FAK and the active forms of M-CSFR and Src were redistributed to the cytoskeleton, where active ERK, JNK and PI3K were detectable. Immunofluorescence showed the presence of FAK and its active form in focal complexes following M-CSF treatment. Moreover, cell spreading and adhesion were impaired when FAK tyrosine phosphorylation was abrogated by either transfection with FRNK, a dominant negative form of FAK, or treatment with a number of inhibitors of upstream FAK-activating signals. These results point to a relevant role for FAK in the regulation of cell spreading and adhesion in macrophages.

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