Filamin depletion blocks endoplasmic spreading and destabilizes force-bearing adhesions

Lynch, Christopher D.; Gauthier, Nils C.; Biais, Nicolas; Lazar, Andre M.; Roca‐Cusachs, Pere; Yu, Cheng-han; Sheetz, Michael P.

doi:10.1091/mbc.e10-08-0661

Cited by 62 publications

(80 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The subsequent build-up of mechanical tension can then relieve the autoinhibition of the other domains promoting strong anchoring. The necessity of multiple parallel bonds for stable anchoring is in accord with the idea that the multiple binding sites of filamin can also induce clustering of transmembrane receptors (10,25). Alternatively, filamin interactions may be substituted or stabilized by other adaptor molecules.…”

Section: Discussionmentioning

confidence: 52%

“…Two prominent examples for interaction partners are integrins (4)(5)(6) or glycoprotein Ib (GPIb), a constituent of a transmembrane complex found in platelets that binds to von Willebrand factor during blood clotting (7)(8)(9). Both the filamin-integrin and the filaminGPIb interactions have been shown to be exposed to mechanical forces in living cells (10,11). The filamin monomer consists of an N-terminal actin-binding domain and a sequence of 24 Ig domain repeats that form the filamin rod (Fig.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Dynamic force sensing of filamin revealed in single-molecule experiments

Rognoni

Stigler

Pelz

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

149

160

View full text Add to dashboard Cite

Mechanical forces are important signals for cell response and development, but detailed molecular mechanisms of force sensing are largely unexplored. The cytoskeletal protein filamin is a key connecting element between the cytoskeleton and transmembrane complexes such as integrins or the von Willebrand receptor glycoprotein Ib. Here, we show using single-molecule mechanical measurements that the recently reported Ig domain pair 20-21 of human filamin A acts as an autoinhibited force-activatable mechanosensor. We developed a mechanical single-molecule competition assay that allows online observation of binding events of target peptides in solution to the strained domain pair. We find that filamin force sensing is a highly dynamic process occurring in rapid equilibrium that increases the affinity to the target peptides by up to a factor of 17 between 2 and 5 pN. The equilibrium mechanism we find here can offer a general scheme for cellular force sensing.optical tweezers | mechanosensing

show abstract

Section: Discussionmentioning

confidence: 52%

mentioning

confidence: 99%

Dynamic force sensing of filamin revealed in single-molecule experiments

Rognoni

Stigler

Pelz

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

149

160

View full text Add to dashboard Cite

show abstract

“…S7B). To check whether filamin A is involved in the formation of the perinuclear actin rim, we used filamin A KO mouse embryonic fibroblast cells (33). However, even in filamin A KO cells, the perinuclear actin rim assembly was still observed after stimulation with A23187 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Mechanical stimulation induces formin-dependent assembly of a perinuclear actin rim

Shao

Mogilner

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

126

129

View full text Add to dashboard Cite

Cells constantly sense and respond to mechanical signals by reorganizing their actin cytoskeleton. Although a number of studies have explored the effects of mechanical stimuli on actin dynamics, the immediate response of actin after force application has not been studied. We designed a method to monitor the spatiotemporal reorganization of actin after cell stimulation by local force application. We found that force could induce transient actin accumulation in the perinuclear region within ∼2 min. This actin reorganization was triggered by an intracellular Ca 2+ burst induced by force application. Treatment with the calcium ionophore A23187 recapitulated the force-induced perinuclear actin remodeling. Blocking of actin polymerization abolished this process. Overexpression of Klarsicht, ANC-1, Syne Homology (KASH) domain to displace nesprins from the nuclear envelope did not abolish Ca 2+ -dependent perinuclear actin assembly. However, the endoplasmic reticulum-and nuclear membrane-associated inverted formin-2 (INF2), a potent actin polymerization activator (mutations of which are associated with several genetic diseases), was found to be important for perinuclear actin assembly. The perinuclear actin rim structure colocalized with INF2 on stimulation, and INF2 depletion resulted in attenuation of the rim formation. Our study suggests that cells can respond rapidly to external force by remodeling perinuclear actin in a unique Ca 2+ -and INF2-dependent manner.force | mechanotransduction | calcium | formin | perinuclear actin rim C ells can sense and adapt to their physical microenvironment through specific mechanosensing mechanisms. These properties are often mediated by the actin cytoskeleton, which can be modulated by a wide range of forces. Fluid shear stress, for example, induces actin stress fiber assembly and realignment along the direction of flow (1-4), whereas the cyclic stretch of an elastic substrate induces a reorientation of stress fibers under some angle to the direction of stretch (5-8). Applying mechanical force to cells by a microneedle results in focal adhesion growth and activation of formin-type actin nucleators (9, 10). Similarly, local application of force through fibronectin or collagen-coated beads trapped by optical or magnetic tweezers leads to the local reorganization of the actin cytoskeleton. This response is associated with reinforcement of bead attachment (11), recruitment of additional actin-associated proteins (12), and activation of a variety of signaling pathways (13)(14)(15)(16)(17). Most studies to date have explored the effects of force on actin structures directly associated with the sites of force application, such as focal adhesions and stress fibers. However, the immediate effect of force on the assembly of actin structures distal from the sites of force application has not been assessed. Such process is despite distal effects having potential implications in the transduction of local forces from the cell periphery to nuclear events (18).In this study, we used a local mecha...

show abstract

“…This was further supported by indications that FLNa-deficiency impaired neuronal migration (Fox and Walsh, 1999;Nagano et al, 2002). Further studies based on knockout mice (Hart et al, 2006;Feng et al, 2006) and the use of small interference RNA showed that while in many settings loss of only one FLN isoform is not sufficient to impair migration, loss of two or more FLNs impaired initiation of migration and cell spreading Lynch et al, 2011;Heuzé et al, 2008). Surprisingly, two recent studies found that FLNa expression negatively correlates with the ability of breast cancer cells to invade and disseminate (Xu et al, 2010;Caruso and Stemmer, 2011).…”

Section: Introductionmentioning

confidence: 93%

Filamin A controls matrix metalloprotease activity and regulates cell invasion in human fibrosarcoma cells.

Baldassarre

Razinia

Brahme

et al. 2012

Journal of Cell Science

View full text Add to dashboard Cite

SummaryFilamins are an important family of actin-binding proteins that, in addition to bundling actin filaments, link cell surface adhesion proteins, signaling receptors and channels to the actin cytoskeleton, and serve as scaffolds for an array of intracellular signaling proteins. Filamins are known to regulate the actin cytoskeleton, act as mechanosensors that modulate tissue responses to matrix density, control cell motility and inhibit activation of integrin adhesion receptors. In this study, we extend the repertoire of filamin activities to include control of extracellular matrix (ECM) degradation. We show that knockdown of filamin increases matrix metalloproteinase (MMP) activity and induces MMP2 activation, enhancing the ability of cells to remodel the ECM and increasing their invasive potential, without significantly altering two-dimensional random cell migration. We further show that within filamin A, the actin-binding domain is necessary, but not sufficient, to suppress the ECM degradation seen in filamin-A-knockdown cells and that dimerization and integrin binding are not required. Filamin mutations are associated with neuronal migration disorders and a range of congenital malformations characterized by skeletal dysplasia and various combinations of cardiac, craniofacial and intestinal anomalies. Furthermore, in breast cancers loss of filamin A has been correlated with increased metastatic potential. Our data suggest that effects on ECM remodeling and cell invasion should be considered when attempting to provide cellular explanations for the physiological and pathological effects of altered filamin expression or filamin mutations.

show abstract

Filamin depletion blocks endoplasmic spreading and destabilizes force-bearing adhesions

Cited by 62 publications

References 45 publications

Dynamic force sensing of filamin revealed in single-molecule experiments

Dynamic force sensing of filamin revealed in single-molecule experiments

Mechanical stimulation induces formin-dependent assembly of a perinuclear actin rim

Filamin A controls matrix metalloprotease activity and regulates cell invasion in human fibrosarcoma cells.

Contact Info

Product

Resources

About