scholarly journals Lipid binding promotes oligomerization and focal adhesion activity of vinculin

2014 ◽  
Vol 207 (5) ◽  
pp. 643-656 ◽  
Author(s):  
Krishna Chinthalapudi ◽  
Erumbi S. Rangarajan ◽  
Dipak N. Patil ◽  
Eric M. George ◽  
David T. Brown ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Lipid Binding ◽  
Stress Fibers ◽  
Actin Binding ◽  
Null Mouse ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Growth And Survival ◽  
Adhesion Activity

Adherens junctions (AJs) and focal adhesion (FA) complexes are necessary for cell migration and morphogenesis, and for the development, growth, and survival of all metazoans. Vinculin is an essential regulator of both AJs and FAs, where it provides links to the actin cytoskeleton. Phosphatidylinositol 4,5-bisphosphate (PIP2) affects the functions of many targets, including vinculin. Here we report the crystal structure of vinculin in complex with PIP2, which revealed that PIP2 binding alters vinculin structure to direct higher-order oligomerization and suggests that PIP2 and F-actin binding to vinculin are mutually permissive. Forced expression of PIP2-binding–deficient mutants of vinculin in vinculin-null mouse embryonic fibroblasts revealed that PIP2 binding is necessary for maintaining optimal FAs, for organization of actin stress fibers, and for cell migration and spreading. Finally, photobleaching experiments indicated that PIP2 binding is required for the control of vinculin dynamics and turnover in FAs. Thus, through oligomerization, PIP2 directs a transient vinculin sequestration at FAs that is necessary for proper FA function.

TDAG51 Deficiency Promotes Migration and Proliferation of Mouse Embryonic Fibroblasts.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3941-3941
Author(s):  
Gazi S. Hossain ◽  
Jeffrey G. Dickhout ◽  
Christopher A. McCulloch ◽  
Ji Zhou ◽  
Linda May ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Migration ◽  
Focal Adhesion ◽  
Laser Scanning ◽  
Collagen Gel ◽  
Wild Type ◽  
Mouse Embryonic Fibroblasts ◽  
Inhibitory Antibody ◽  
Embryonic Fibroblasts ◽  
Wild Type Mefs

Abstract T-cell death associated gene 51 (TDAG51) is a member of the pleckstrin homology-related domain family having pro-apoptotic characteristics. Indirect immunofluorescence studies demonstrate that endogenous TDAG51 co-localizes with focal adhesion kinase (FAK), a molecule found within focal adhesion complexes (FACs). FACs are believed to be the structural connection between the extracellular matrix (ECM) and actin cytoskeleton. Given that overexpression of TDAG51 promotes detachment-induced programmed cell death (PCD) and that TDAG51 co-localizes with FAK, it was proposed that deficiency of TDAG51 may stabilize FAC assembly, thereby affecting actin cytoskeletal organization and/or cell migration. To better understand the role of TDAG51 in cell migration, mouse embryonic fibroblasts (MEFs) deficient in TDAG51 were derived from TDAG51-deficient mice and compared to wild type MEFs. TDAG51-deficient MEFs showed increased migration following monolayer disruption or in response to chemotaxis on fibronectin-coated Boyden Chambers. In addition, loss of TDAG51 promotes the proliferation of MEFs. Collagen gel contraction experiments were performed to confirm the aggregate effect of the differences in migration and proliferation observed in TDAG51-deficient MEFs. Interestingly, migration and proliferation can be blocked by active b-integrin inhibitory antibody. In terms of a cellular phenotype, migratory TDAG51-deficient MEFs have distinct filopodial and lamellipodial extentions, compared to migratory wild type MEFs. Laser scanning confocal microscope demonstrated a significant decrease in F-actin stress fibers as well as increased distribution of vinculin within the lamellipodial protrusions of TDAG51-deficient MEFs. Importantly, reintroduction of TDAG51 into TDAG51-deficient MEFs reversed these phenotypic changes. Taken together, our findings suggest that loss of TDAG51 affects cell migration and proliferation through modifying focal adhesion complex assembly and cytoskeletal rearrangements.

scholarly journals Rab40/Cullin5 complex regulates EPLIN and actin cytoskeleton dynamics during cell migration and invasion

2021 ◽  
Author(s):  
Erik S Linklater ◽  
Emily Duncan ◽  
Ke Jun Han ◽  
Algirdas Kaupinis ◽  
Mindaugas Valius ◽  
...  
Keyword(s):  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Leading Edge ◽  
Stress Fibers ◽  
Actin Dynamics ◽  
Migration And Invasion ◽  
Matrix Remodeling ◽  
Cell Migration And Invasion

Rab40b is a SOCS box containing protein that regulates the secretion of MMPs to facilitate extracellular matrix remodeling during cell migration. Here we show that Rab40b interacts with Cullin5 via the Rab40b SOCS domain. We demonstrate that loss of Rab40b/Cullin5 binding decreases cell motility and invasive potential, and show that defective cell migration and invasion stem from alteration to the actin cytoskeleton, leading to decreased invadopodia formation, decreased actin dynamics at the leading edge, and an increase in stress fibers. We also show that these stress fibers anchor at less dynamic, more stable focal adhesions. Mechanistically, changes in the cytoskeleton and focal adhesion dynamics are mediated in part by EPLIN, which we demonstrate to be a binding partner of Rab40b and a target for Rab40b/Cullin5 dependent localized ubiquitylation and degradation. Thus, we propose a model where the Rab40b/Cullin5 dependent ubiquitylation regulates EPLIN localization to promote cell migration and invasion by altering focal adhesion and cytoskeletal dynamics.

Indomethacin Delays Gastric Restitution: Association with the Inhibition of Focal Adhesion Kinase and Tensin Phosphorylation and Reduced Actin Stress Fibers

2002 ◽  
Vol 227 (6) ◽  
pp. 412-424 ◽  
Author(s):  
Imre L. Szabó ◽  
Rama Pai ◽  
Michael K. Jones ◽  
George R. Ehring ◽  
Hirofumi Kawanaka ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Stress Fiber ◽  
Fiber Formation ◽  
Stress Fibers ◽  
Actin Stress Fiber ◽  
Stress Fiber Formation ◽  
Actin Stress Fiber Formation

Repair of superficial gastric mucosal injury is accomplished by the process of restitution—migration of epithelial cells to restore continuity of the mucosal surface. Actin filaments, focal adhesions, and focal adhesion kinase (FAK) play crucial roles in cell motility essential for restitution. We studied whether epidermal growth factor (EGF) and/or indomethacin (IND) affect cell migration, actin stress fiber formation, and/or phosphorylation of FAK and tensin in wounded gastric monolayers. Human gastric epithelial monolayers (MKN 28 cells) were wounded and treated with either vehicle or 0.5 mM IND for 16 hr followed by EGF. EGF treatment significantly stimulated cell migration and actin stress fiber formation, and increased FAK localization to focal adhesions, and phosphorylation of FAK and tensin, whereas IND inhibited all these at the baseline and EGF-stimulated conditions. IND-induced inhibition of FAK phosphorylation preceded changes in actin polymerization, indicating that actin depolymerization might be the consequence of decreased FAK activity. In in vivo experiments, rats received either vehicle or IND (5 mg/kg i.g.), and 3 min later, they received water or 5% hypertonic NaCl; gastric mucosa was obtained at 1, 4, and 8 hr after injury. Four and 8 hr after hypertonic injury, FAK phosphorylation was induced in gastric mucosa compared with controls. IND pretreatment significantly delayed epithelial restitution in vivo, and reduced FAK phosphorylation and recruitment to adhesion points, as well as actin stress fiber formation in migrating surface epithelial cells. Our study indicates that FAK, tensin, and actin stress fibers are likely mediators of EGF-stimulated cell migration in wounded human gastric monolayers and potential targets for IND-induced inhibition of restitution.

UV light induces premature senescence in Akt1-null mouse embryonic fibroblasts by increasing intracellular levels of ROS

2009 ◽  
Vol 383 (3) ◽  
pp. 358-362 ◽  
Author(s):  
Hye Jin Jee ◽  
Hyun-Ju Kim ◽  
Ae Jeong Kim ◽  
Yoe-Sik Bae ◽  
Sun Sik Bae ◽  
...  
Keyword(s):  
Uv Light ◽  
Null Mouse ◽  
Premature Senescence ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts

Differential expression of prohibitin and regulation of apoptosis in wild-type and COX-2 null mouse embryonic fibroblasts

2009 ◽  
Vol 53 (2) ◽  
pp. 157-159 ◽  
Author(s):  
Young-Hoon Kim ◽  
Hyangkyu Lee ◽  
Tae-Yoon Kim ◽  
Hyang-Ran Hwang ◽  
Sang Chul Lee
Keyword(s):  
Differential Expression ◽  
Null Mouse ◽  
Wild Type ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Cox 2

scholarly journals <i>Gsta</i>4 Null Mouse Embryonic Fibroblasts Exhibit Enhanced Sensitivity to Oxidants: Role of 4-Hydroxynonenal in Oxidant Toxicity

2013 ◽  
Vol 02 (01) ◽  
pp. 1-11 ◽  
Author(s):  
Kevin E. McElhanon ◽  
Chhanda Bose ◽  
Rajendra Sharma ◽  
Liping Wu ◽  
Yogesh C. Awasthi ◽  
...  
Keyword(s):  
Null Mouse ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Enhanced Sensitivity
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