scholarly journals Vimentin tunes cell migration on collagen by controlling β1 integrin activation and clustering

2021 ◽  
Vol 134 (6) ◽  
pp. jcs254359 ◽  
Author(s):  
Zofia Ostrowska-Podhorodecka ◽  
Isabel Ding ◽  
Wilson Lee ◽  
Jelena Tanic ◽  
Sevil Abbasi ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesions ◽  
Adaptor Protein ◽  
Mesenchymal Cell ◽  
Β1 Integrin ◽  
Structural Protein ◽  
Integrin Activation ◽  
Vimentin Expression ◽  
Downstream Effector ◽  
Focal Adhesion Proteins

ABSTRACTVimentin is a structural protein that is required for mesenchymal cell migration and directly interacts with actin, β1 integrin and paxillin. We examined how these interactions enable vimentin to regulate cell migration on collagen. In fibroblasts, depletion of vimentin increased talin-dependent activation of β1 integrin by more than 2-fold. Loss of vimentin was associated with reduction of β1 integrin clustering by 50% and inhibition of paxillin recruitment to focal adhesions by more than 60%, which was restored by vimentin expression. This reduction of paxillin was associated with 65% lower Cdc42 activation, a 60% reduction of cell extension formation and a greater than 35% decrease in cell migration on collagen. The activation of PAK1, a downstream effector of Cdc42, was required for vimentin phosphorylation and filament maturation. We propose that vimentin tunes cell migration through collagen by acting as an adaptor protein for focal adhesion proteins, thereby regulating β1 integrin activation, resulting in well-organized, mature integrin clusters.This article has an associated First Person interview with the first author of the paper.

scholarly journals Different translation dynamics of β-and γ-actin regulates cell migration

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Pavan Vedula ◽  
Satoshi Kurosaka ◽  
Brittany MacTaggart ◽  
Qin Ni ◽  
Garegin Papoian ◽  
...  
Keyword(s):  
Cell Migration ◽  
Amino Acid ◽  
Single Molecule ◽  
Amino Acid Level ◽  
Focal Adhesions ◽  
Mesenchymal Cell ◽  
Cell Periphery ◽  
Pronounced Difference ◽  
Coding Sequence

β- and γ-cytoplasmic actins are ubiquitously expressed in every cell type and are nearly identical at the amino acid level but play vastly different roles in vivo. Their essential roles in embryogenesis and mesenchymal cell migration critically depend on the nucleotide sequences of their genes, rather than their amino acid sequence, however it is unclear which gene elements underlie this effect. Here we address the specific role of the coding sequence in β- and γ-cytoplasmic actins' intracellular functions, using stable polyclonal populations of immortalized mouse embryonic fibroblasts with exogenously expressed actin isoforms and their 'codon-switched' variants. When targeted to the cell periphery using the β-actin 3′UTR, β-actin and γ-actin have differential effects on cell migration. These effects directly depend on the coding sequence. Single molecule measurements of actin isoform translation, combined with fluorescence recovery after photobleaching, demonstrate a pronounced difference in β- and γ-actins' translation elongation rates in cells, leading to changes in their dynamics at the focal adhesions, impairments in actin bundle formation, and reduced cell anchoring to the substrate during migration. Our results demonstrate that coding sequence-mediated differences in actin translation play a key role in cell migration.

scholarly journals Mena binds α5 integrin directly and modulates α5β1 function

2012 ◽  
Vol 198 (4) ◽  
pp. 657-676 ◽  
Author(s):  
Stephanie L. Gupton ◽  
Daisy Riquelme ◽  
Shannon K. Hughes-Alford ◽  
Jenny Tadros ◽  
Shireen S. Rudina ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Adhesion ◽  
Growth Factor Receptor ◽  
Focal Adhesions ◽  
Fibronectin Receptor ◽  
Focal Adhesion Proteins ◽  
Carboxy Terminal ◽  
Bidirectional Interactions ◽  
Dependent Processes ◽  
Α5β1 Integrin

Mena is an Ena/VASP family actin regulator with roles in cell migration, chemotaxis, cell–cell adhesion, tumor cell invasion, and metastasis. Although enriched in focal adhesions, Mena has no established function within these structures. We find that Mena forms an adhesion-regulated complex with α5β1 integrin, a fibronectin receptor involved in cell adhesion, motility, fibronectin fibrillogenesis, signaling, and growth factor receptor trafficking. Mena bound directly to the carboxy-terminal portion of the α5 cytoplasmic tail via a 91-residue region containing 13 five-residue “LERER” repeats. In fibroblasts, the Mena–α5 complex was required for “outside-in” α5β1 functions, including normal phosphorylation of FAK and paxillin and formation of fibrillar adhesions. It also supported fibrillogenesis and cell spreading and controlled cell migration speed. Thus, fibroblasts require Mena for multiple α5β1-dependent processes involving bidirectional interactions between the extracellular matrix and cytoplasmic focal adhesion proteins.

scholarly journals Akt Phosphorylation of Serine 21 on Pak1 Modulates Nck Binding and Cell Migration

2003 ◽  
Vol 23 (22) ◽  
pp. 8058-8069 ◽  
Author(s):  
Guo-Lei Zhou ◽  
Ya Zhuo ◽  
Charles C. King ◽  
Benjamin H. Fryer ◽  
Gary M. Bokoch ◽  
...  
Keyword(s):  
Cell Migration ◽  
Protein Kinases ◽  
Cellular Proliferation ◽  
Fluorescent Protein ◽  
Focal Adhesions ◽  
Adaptor Protein ◽  
Small Gtpases ◽  
Akt Phosphorylation ◽  
Green Fluorescent

ABSTRACT The p21-activated protein kinases (Paks) regulate cellular proliferation, differentiation, transformation, and survival through multiple downstream signals. Paks are activated directly by the small GTPases Rac and Cdc42 and several protein kinases including Akt and PDK-1. We found that Akt phosphorylated and modestly activated Pak1 in vitro. The major site phosphorylated by Akt on Pak1 mapped to serine 21, a site originally shown to be weakly autophosphorylated on Pak1 when Cdc42 or Rac activates it. A peptide derived from the region surrounding serine 21 was a substrate for Akt but not Pak1 in vitro, and Akt stimulated serine 21 phosphorylation on the full-length Pak1 much better than Rac did. The adaptor protein Nck binds Pak near serine 21, and its association is regulated by phosphorylation of this site. We found that either treatment of Pak1 in vitro with Akt or coexpression of constitutively active Akt with Pak1 reduced Nck binding to Pak1. In HeLa cells, green fluorescent protein-tagged Pak1 was concentrated at focal adhesions and was released when Akt was cotransfected. A peptide containing the Nck binding site of Pak1 fused to a portion of human immunodeficiency virus Tat to allow it to enter cells was used to test the functional importance of Nck/Pak binding in Akt-stimulated cell migration. This Tat-Nck peptide reduced Akt-stimulated cell migration. Together, these data suggest that Akt modulates the association of Pak with Nck to regulate cell migration.

scholarly journals Semaphorin 4D/Plexin-B1–mediated R-Ras GAP activity inhibits cell migration by regulating β1 integrin activity

2006 ◽  
Vol 173 (4) ◽  
pp. 601-613 ◽  
Author(s):  
Izumi Oinuma ◽  
Hironori Katoh ◽  
Manabu Negishi
Keyword(s):  
Cell Migration ◽  
Cell Types ◽  
Β1 Integrin ◽  
Integrin Activation ◽  
Kinase Activation ◽  
Surface Receptors ◽  
Semaphorin 4D ◽  
Ras Activation ◽  
Ras Family

Plexins are cell surface receptors for semaphorins and regulate cell migration in many cell types. We recently reported that the semaphorin 4D (Sema4D) receptor Plexin-B1 functions as a GTPase-activating protein (GAP) for R-Ras, a member of Ras family GTPases implicated in regulation of integrin activity and cell migration (Oinuma, I., Y. Ishikawa, H. Katoh, and M. Negishi. 2004. Science. 305:862–865). We characterized the role of R-Ras downstream of Sema4D/Plexin-B1 in cell migration. Activation of Plexin-B1 by Sema4D suppressed the ECM-dependent R-Ras activation, R-Ras–mediated phosphatydylinositol 3-kinase activation, and β1 integrin activation through its R-Ras GAP domain, leading to inhibition of cell migration. In addition, inactivation of R-Ras by overexpression of the R-Ras–specific GAP or knockdown of R-Ras by RNA interference was sufficient for suppressing β1 integrin activation and cell migration in response to the ECM stimulation. Thus, we conclude that R-Ras activity is critical for ECM-mediated β1 integrin activation and cell migration and that inactivation of R-Ras by Sema4D/Plexin-B1–mediated R-Ras GAP activity controls cell migration by modulating the activity of β1 integrins.

scholarly journals RhoB regulates cell migration through altered focal adhesion dynamics

Open Biology ◽  
2012 ◽  
Vol 2 (5) ◽  
pp. 120076 ◽  
Author(s):  
Francisco M. Vega ◽  
Audrey Colomba ◽  
Nicolas Reymond ◽  
Mairian Thomas ◽  
Anne J. Ridley
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Rho Gtpase ◽  
Focal Adhesions ◽  
Β1 Integrin ◽  
Membrane Ruffling ◽  
Similar Phenotype ◽  
And Migration ◽  
Directional Cell Migration ◽  
Chemotactic Gradient

The Rho GTPase RhoB has been shown to affect cell migration, but how it does this is not clear. Here we show that cells depleted of RhoB by RNAi are rounded and have defects in Rac-mediated spreading and lamellipodium extension, although they have active membrane ruffling around the periphery. Depletion of the exchange factor GEF-H1 induces a similar phenotype. RhoB-depleted cells migrate faster, but less persistently in a chemotactic gradient, and frequently round up during migration. RhoB-depleted cells have similar numbers of focal adhesions to control cells during spreading and migration, but show more diffuse and patchy contact with the substratum. They have lower levels of surface β1 integrin, and β1 integrin activity is reduced in actin-rich protrusions. We propose that RhoB contributes to directional cell migration by regulating β1 integrin surface levels and activity, thereby stabilizing lamellipodial protrusions.

scholarly journals Retrograde Fluxes of Focal Adhesion Proteins in Response to Cell Migration and Mechanical Signals

2007 ◽  
Vol 18 (11) ◽  
pp. 4519-4527 ◽  
Author(s):  
Wei-hui Guo ◽  
Yu-li Wang
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Mechanical Load ◽  
Focal Adhesions ◽  
Force Production ◽  
Retrograde Transport ◽  
Mechanical Stimuli ◽  
Adhesion Proteins ◽  
Mechanical Signals ◽  
Focal Adhesion Proteins

Recent studies suggest that mechanical signals mediated by the extracellular matrix play an essential role in various physiological and pathological processes; yet, how cells respond to mechanical stimuli remains elusive. Using live cell fluorescence imaging, we found that actin filaments, in association with a number of focal adhesion proteins, including zyxin and vasodilator-stimulated phosphoprotein, undergo retrograde fluxes at focal adhesions in the lamella region. This flux is inversely related to cell migration, such that it is amplified in fibroblasts immobilized on micropatterned islands. In addition, the flux is regulated by mechanical signals, including stretching forces applied to flexible substrates and substrate stiffness. Conditions favoring the flux share the common feature of causing large retrograde displacements of the interior actin cytoskeleton relative to the substrate anchorage site, which may function as a switch translating mechanical input into chemical signals, such as tyrosine phosphorylation. In turn, the stimulation of actin flux at focal adhesions may function as part of a feedback mechanism, regulating structural assembly and force production in relation to cell migration and mechanical load. The retrograde transport of associated focal adhesion proteins may play additional roles in delivering signals from focal adhesions to the interior of the cell.

scholarly journals Migration regulates cellular mechanical states

2019 ◽  
Vol 30 (26) ◽  
pp. 3104-3111 ◽  
Author(s):  
Stephanie S. Chang ◽  
Andrew D. Rape ◽  
Stephanie A. Wong ◽  
Wei-hui Guo ◽  
Yu-li Wang
Keyword(s):  
Cell Migration ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Traction Forces ◽  
Adhesion Proteins ◽  
Focal Adhesion Proteins

Cell migration has a profound effect on the generation of traction forces and the phosphorylation of focal adhesion proteins. The mechanism may involve the dynamic turnover of focal adhesions during cell migration and mechanical interactions between nascent and preexisting focal adhesions.

scholarly journals Actin stress fibre subtypes in mesenchymal-migrating cells

Open Biology ◽  
2013 ◽  
Vol 3 (6) ◽  
pp. 130001 ◽  
Author(s):  
Tea Vallenius
Keyword(s):  
Cell Migration ◽  
Myosin Ii ◽  
Focal Adhesions ◽  
Mesenchymal Cell ◽  
Molecular Composition ◽  
Current View ◽  
Stress Fibre ◽  
Actin Stress Fibre ◽  
Extracellular Matrix Remodelling ◽  
Migrating Cells

Mesenchymal cell migration is important for embryogenesis and tissue regeneration. In addition, it has been implicated in pathological conditions such as the dissemination of cancer cells. A characteristic of mesenchymal-migrating cells is the presence of actin stress fibres, which are thought to mediate myosin II-based contractility in close cooperation with associated focal adhesions. Myosin II-based contractility regulates various cellular activities, which occur in a spatial and temporal manner to achieve directional cell migration. These myosin II-based activities involve the maturation of integrin-based adhesions, generation of traction forces, establishment of the front-to-back polarity axis, retraction of the trailing edge, extracellular matrix remodelling and mechanotransduction. Growing evidence suggests that actin stress fibre subtypes, namely dorsal stress fibres, transverse arcs and ventral stress fibres, could provide this spatial and temporal myosin II-based activity. Consistent with their functional differences, recent studies have demonstrated that the molecular composition of actin stress fibre subtypes differ significantly. This present review focuses on the current view of the molecular composition of actin stress fibre subtypes and how these fibre subtypes regulate mesenchymal cell migration.

scholarly journals Calpain-mediated Proteolysis of Paxillin Negatively Regulates Focal Adhesion Dynamics and Cell Migration

2011 ◽  
Vol 286 (12) ◽  
pp. 9998-10006 ◽  
Author(s):  
Christa L. Cortesio ◽  
Lindsy R. Boateng ◽  
Timothy M. Piazza ◽  
David A. Bennin ◽  
Anna Huttenlocher
Keyword(s):  
Cell Migration ◽  
Cell Motility ◽  
Focal Adhesion ◽  
Focal Adhesions ◽  
Adaptor Protein ◽  
Time Lapse ◽  
Negative Regulator ◽  
Proteolytic Fragment ◽  
Proteolytic Site ◽  
And Migration

The dynamic turnover of integrin-mediated adhesions is important for cell migration. Paxillin is an adaptor protein that localizes to focal adhesions and has been implicated in cell motility. We previously reported that calpain-mediated proteolysis of talin1 and focal adhesion kinase mediates adhesion disassembly in motile cells. To determine whether calpain-mediated paxillin proteolysis regulates focal adhesion dynamics and cell motility, we mapped the preferred calpain proteolytic site in paxillin. The cleavage site is between the paxillin LD1 and LD2 motifs and generates a C-terminal fragment that is similar in size to the alternative product paxillin delta. The calpain-generated proteolytic fragment, like paxillin delta, functions as a paxillin antagonist and impairs focal adhesion disassembly and migration. We generated mutant paxillin with a point mutation (S95G) that renders it partially resistant to calpain proteolysis. Paxillin-deficient cells that express paxillin S95G display increased turnover of zyxin-containing adhesions using time-lapse microscopy and also show increased migration. Moreover, cancer-associated somatic mutations in paxillin are common in the N-terminal region between the LD1 and LD2 motifs and confer partial calpain resistance. Taken together, these findings suggest a novel role for calpain-mediated proteolysis of paxillin as a negative regulator of focal adhesion dynamics and migration that may function to limit cancer cell invasion.

scholarly journals Crk adaptor protein-induced phosphorylation of Gab1 on tyrosine 307 via Src is important for organization of focal adhesions and enhanced cell migration

Cell Research ◽  
2009 ◽  
Vol 19 (5) ◽  
pp. 638-650 ◽  
Author(s):  
Takuya Watanabe ◽  
Masumi Tsuda ◽  
Yoshinori Makino ◽  
Tassos Konstantinou ◽  
Hiroshi Nishihara ◽  
...  
Keyword(s):  
Cell Migration ◽  
Focal Adhesions ◽  
Adaptor Protein
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