The last but not the least: The origin and significance of trailing adhesions in fibroblastic cells

Rid, Raphaela; Schiefermeier, Natalia; Grigoriev, Ilya; Small, J. Victor; Kaverina, Irina

doi:10.1002/cm.20076

Cited by 65 publications

(69 citation statements)

References 35 publications

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“…The relationship between the number of focal contacts and the cell movement, speed and directionality shown in Fig. 7 concurs with much earlier data showing that cells slow down as the number of focal contacts [56][57][58] and directionality of their movement increases [59,60]. Indeed, this is already evident in calreticulin KO cells, which still have some albeit much less numerous stress fibers and focal contacts.…”

Section: Functional Consequences Of Irs-1 Inhibitionsupporting

confidence: 89%

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Czarnowski

Papp

Száraz

et al. 2014

Cellular and Molecular Biology Letters

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Cellular adhesion to the underlying substratum is regulated through numerous signaling pathways. It has been suggested that insulin receptor substrate 1 (IRS-1) is involved in some of these pathways, via association with and activation of transmembrane integrins. Calreticulin, as an important endoplasmic reticulum-resident, calcium-binding protein with a chaperone function, plays an obvious role in proteomic expression. Our previous work showed that calreticulin mediates cell adhesion not only by affecting protein expression but also by affecting the state of regulatory protein phosphorylation, such as that of c-src. Here, we demonstrate that calreticulin affects the abundance of IRS-1 such that the absence of calreticulin is paralleled by a decrease in IRS-1 levels and the unregulated overexpression of calreticulin is accompanied by an increase in IRS-1 levels. These changes in the abundance of calreticulin and IRS-1 are accompanied by changes in cell-substratum adhesiveness and phosphorylation, such that increases in the expression of calreticulin and IRS-1 are paralleled by an increase in focal contact-based cellsubstratum adhesiveness, and a decrease in the expression of these proteins brings about a decrease in cell-substratum adhesiveness. Wild type and calreticulin-null mouse embryonic fibroblasts (MEFs) were cultured and the IRS-1 isoform profile was assessed. Differences in morphology and motility were also quantified. While no substantial differences in the speed of locomotion were found, the directionality of cell movement was greatly promoted by the presence of calreticulin. Calreticulin expression was also found to have a dramatic effect on the phosphorylation state of serine 636 of IRS-1, such that phosphorylation of IRS-1 on serine 636 increased radically in the absence of calreticulin. Most importantly, treatment of cells with the RhoA/ROCK inhibitor, Y-27632, which among its many effects also inhibited serine 636 phosphorylation of IRS-1, had profound effects on cell-substratum adhesion, in that it suppressed focal contacts, induced extensive close contacts, and increased the strength of adhesion. The latter effect, while counterintuitive, can be explained by the close contacts comprising labile bonds but in large numbers. In addition, the lability of bonds in close contacts would permit fast locomotion. An interesting and novel finding is that Y-27632 treatment of MEFs releases them from contact inhibition of locomotion, as evidenced by the invasion of a cell's underside by the thin lamellae and filopodia of a cell in close apposition.

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Section: Functional Consequences Of Irs-1 Inhibitionsupporting

confidence: 89%

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Czarnowski

Papp

Száraz

et al. 2014

Cellular and Molecular Biology Letters

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“…Another possibility is that normal levels and regulation of FAK-associated Src signaling promote adhesion turnover, while enhanced deregulated signaling achieved by the FAK/Src chimera has the opposite effect of promoting assembly. A third consideration is that ULPA resemble the class of trailing "sliding adhesions" that form at the cell rear and flanks and mature rapidly into larger stress fiber-associated structures [Rid et al, 2005]. Sliding adhesions are distinct from the stationary adhesions studied by Webb et al and could be subject to different regulation.…”

Section: Discussionmentioning

confidence: 99%

“…Both p130Cas [Brábek et al, 2004] and the Arp2/3 complex ] are known to have roles in podosome assembly. Since adhesion formation is associated with intensive actin polymerization [Rid et al, 2005], the rapid assembly of new adhesions and ultimate ULPA formation may also be a consequence of FAK/Src signaling driving actin polymerization.…”

Section: Discussionmentioning

confidence: 99%

A FAK/Src chimera with gain‐of‐function properties promotes formation of large peripheral adhesions associated with dynamic actin assembly

Siesser

Meenderink

Ryzhova

et al. 2007

Cell Motil. Cytoskeleton

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Formation of a complex between the tyrosine kinases FAK and Src is a key integrin-mediated signaling event implicated in cell motility, survival, and proliferation. Past studies indicate that FAK functions in the complex primarily as a "scaffold," acting to recruit and activate Src within cell/matrix adhesions. To study the cellular impact of FAK-associated Src signaling we developed a novel gainof-function approach that involves expressing a chimeric protein with the FAK kinase domain replaced by the Src kinase domain. This FAK/Src chimera is subject to adhesion-dependent activation and promotes tyrosine phosphorylation of p130Cas and paxillin to higher steady-state levels than is achieved by wild-type FAK. When expressed in FAK -/-mouse embryo fibroblasts, the FAK/Src chimera resulted in a striking cellular phenotype characterized by unusual large peripheral adhesions, enhanced adhesive strength, and greatly reduced motility. Live cell imaging of the chimeraexpressing FAK -/-cells provided evidence that the large peripheral adhesions are associated with a dynamic actin assembly process that is sensitive to a Src-selective inhibitor. These findings suggest that FAK-associated Src kinase activity has the capacity to promote adhesion integrity and actin assembly.

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“…As the lamellipodia advances and the trailing edge stretches the cell becomes elongated. Inward movement and subsequent release of adhesions at the rear leads to trailingedge retraction and cell body advance whereupon the cell becomes more rounded (symmetrical) and the cycle is repeated (Rid et al, 2005;Smilenov et al, 1999;Webb et al, 2005). Fibroblast movement requires spatial regulation of cell attachment to extra cellular matrix (ECM).…”

Section: Introductionmentioning

confidence: 99%

“…However, the trailing edge of the cells must be able to detach from the substrate to permit cell translocation (Regen and Horwitz, 1992). Detachment of the trailing edge requires continued myosin-driven cytoskeletal tension that results in movement and ultimate release (disassociation/ degradation) of adhesions within the trailing edge (Palecek et al, 1998;Palecek et al, 1996;Rid et al, 2005;Smilenov et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

FAK, PDZ-RhoGEF and ROCKII cooperate to regulate adhesion movement and trailing-edge retraction in fibroblasts

Iwanicki

Vomastek

Tilghman

et al. 2008

Journal of Cell Science

107

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A key step in cell migration is the dynamic formation and disassembly of adhesions at the front and the concomitant movement and release of adhesions in the rear of the cell. Fibroblasts maintained in the absence of serum have stable adhesions within the rear of the cell and exhibit reduced trailing-edge retraction resulting in an elongated cell phenotype. Addition of lysophosphatidic acid (LPA) induced the movement of adhesions and retraction of the trailing edge, thus mimicking tail retraction in a migrating cell. Focal adhesion kinase (FAK), guanine nucleotide exchange factors (GEF) for Rho and the Rho effector Rho kinase II (ROCKII) are crucial for the regulation of adhesion movement and trailing-edge retraction. Downregulation of FAK by small interfering RNAs or small hairpin RNAs blocked LPA-induced adhesion movement and restoration of cell shape. This phenotype was rescued by the ectopic expression of PDZ-RhoGEF or a RhoA-effector-domain mutant that activates ROCK. Knockdown of PDZ-RhoGEF or ROCKII inhibited LPA-induced trailing-edge retraction and adhesion movement. Moreover, overexpressed PDZ-RhoGEF co-immunoprecipitated with FAK and localized to FAK-containing adhesions. These studies support a model in which FAK and PDZ-RhoGEF cooperate to induce Rho/ROCKII-dependent focal adhesion movement and trailing-edge retraction in response to LPA.

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The last but not the least: The origin and significance of trailing adhesions in fibroblastic cells

Cited by 65 publications

References 35 publications

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

A FAK/Src chimera with gain‐of‐function properties promotes formation of large peripheral adhesions associated with dynamic actin assembly

FAK, PDZ-RhoGEF and ROCKII cooperate to regulate adhesion movement and trailing-edge retraction in fibroblasts

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