The residence time of focal adhesion kinase (FAK) and paxillin at focal adhesions in renal epithelial cells is determined by adhesion size, strength and life cycle status

Campylobacter jejuni is a major foodborne pathogen that causes severe gastritis. We investigated the dynamics of focal adhesion structure and function in C. jejuni -infected epithelial cells.

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Oxazolone-induced over-expression of focal adhesion kinase in colonic epithelial cells of colitis mouse model

FEBS Letters ◽

10.1016/j.febslet.2010.07.054 ◽

2010 ◽

Vol 584 (18) ◽

pp. 3949-3954 ◽

Cited By ~ 11

Author(s):

Yanju Ma ◽

Shingo Semba ◽

Atsuo Maemoto ◽

Masayuki Takeuchi ◽

Isamu Kameshita ◽

...

Keyword(s):

Epithelial Cells ◽

Mouse Model ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Colonic Epithelial Cells ◽

Over Expression

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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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Src and focal adhesion kinase mediate mechanical strain-induced proliferation and ERK1/2 phosphorylation in human H441 pulmonary epithelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00529.2006 ◽

2007 ◽

Vol 292 (5) ◽

pp. C1701-C1713 ◽

Cited By ~ 59

Author(s):

Lakshmi S. Chaturvedi ◽

Harold M. Marsh ◽

Marc D. Basson

Keyword(s):

Epithelial Cells ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Mechanical Strain ◽

A549 Cells ◽

Cyclic Strain ◽

Short Interfering Rna ◽

Erk Activation ◽

Pulmonary Epithelial Cells ◽

Interfering Rna

Pulmonary epithelial cells are exposed to repetitive deformation during physiological breathing and mechanical ventilation. Such deformation may influence pulmonary growth, development, and barotrauma. Although deformation stimulates proliferation and activates extracellular signal-regulated kinases (ERK1/2) in human pulmonary epithelial H441 cells, the upstream mechanosensors that induce ERK activation are poorly understood. We investigated whether c-Src or focal adhesion kinase (FAK) mediates cyclic mechanical strain-induced ERK1/2 activation and proliferation in human pulmonary epithelial (NCI-H441) cells. The H441 and A549 cells were grown on collagen I-precoated membranes and were subjected to an average 10% cyclic mechanical strain at 20 cycles/min. Cyclic strain activated Src within 2 min by increasing phosphorylation at Tyr418, followed by rapid phosphorylation of FAK at Tyr397 and Tyr576 and ERK1/2 at Thr202/Tyr204 ( n = 5, P < 0.05). Twenty-four (A549 cells) and 24–72 h (H441 cells) of cyclic mechanical strain increased cell numbers compared with static culture. Twenty-four hours of cyclic strain also increased H441 FAK, Src, and ERK phosphorylation without affecting total FAK, Src, or ERK protein. The mitogenic effect was blocked by Src (10 μmol/l PP2 or short interfering RNA targeted to Src) or MEK (50 μmol/l PD-98059) inhibition. PP2 also blocked strain-induced phosphorylation of FAK-Tyr576 and ERK-Thr202/Tyr204 but not FAK-Tyr397. Reducing FAK by FAK-targeted short interfering RNA blocked mechanical strain-induced mitogenicity and significantly attenuated strain-induced ERK activation but not strain-induced Src phosphorylation. Together, these results suggest that repetitive mechanical deformation induced by ventilation supports pulmonary epithelial proliferation by a pathway involving Src, FAK, and then ERK signaling.

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

Journal of Cellular Physiology ◽

10.1002/jcp.22084 ◽

2010 ◽

pp. n/a-n/a ◽

Cited By ~ 14

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

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Autophosphorylation of Tyr397 and its phosphorylation by Src-family kinases are altered in focal-adhesion-kinase neuronal isoforms

Biochemical Journal ◽

10.1042/bj3480119 ◽

2000 ◽

Vol 348 (1) ◽

pp. 119-128 ◽

Cited By ~ 16

Author(s):

Madeleine TOUTANT ◽

Jeanne-Marie STUDLER ◽

Ferran BURGAYA ◽

Alicia COSTA ◽

Pascal EZAN ◽

...

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Fluorescent Protein ◽

Focal Adhesions ◽

Src Family Kinases ◽

Critical Residue ◽

Green Fluorescent ◽

And Function

In brain, focal adhesion kinase (FAK) is regulated by neurotransmitters and has a higher molecular mass than in other tissues, due to alternative splicing. Two exons code for additional peptides of six and seven residues (‘boxes’ 6 and 7), located on either side of Tyr397, which increase its autophosphorylation. Using in situ hybridization and a monoclonal antibody (Mab77) which does not recognize FAK containing box 7, we show that, although mRNAs coding for boxes 6 and 7 have different patterns of expression in brain, FAK+6,7 is the main isoform in forebrain neurons. The various FAK isoforms fused to green fluorescent protein were all targeted to focal adhesions in non-neuronal cells. Phosphorylation-state-specific antibodies were used to study in detail the phosphorylation of Tyr397, a critical residue for the activation and function of FAK. The presence of boxes 6 and 7 increased autophosphorylation of Tyr397 independently and additively, whereas they had a weak effect on FAK kinase activity towards poly(Glu,Tyr). Src-family kinases were also able to phosphorylate Tyr397 in cells, but this phosphorylation was decreased in the presence of box 6 or 7, and abolished in the presence of both. Thus the additional exons characteristic of neuronal isoforms of FAK do not alter its targeting, but change dramatically the phosphorylation of Tyr397. They increase its autophosphorylation in vitro and in transfected COS-7 cells, whereas they prevent its phosphorylation when co-transfected with Src-family kinases.

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