The Structural Basis of Actin Organization by Vinculin and Metavinculin

Kim, Laura Y.; Thompson, Peter M.; Lee, Hyunna T.; Pershad, Mihir; Campbell, Sharon L.; Alushin, Gregory M.

doi:10.1016/j.jmb.2015.09.031

Cited by 51 publications

(116 citation statements)

References 72 publications

Supporting

Mentioning

104

Contrasting

Order By: Relevance

“…The IA and VA mutations reduced the coprecipitation of vinculin to 42% and 53%, respectively, as previously reported [18, 22]. Given that the vinculin tail domain has two actin-binding surfaces [18–20], these results suggest that the IA and VA mutations perturb one of the actin-binding surfaces, but the other binding surface may still bind actin. Although IA or VA mutation did not affect the binding of VBS3(talin)-activated vinculin to actin in vitro , these mutation are known to have effects on vinculin behaviors in cells [18, 22], indicating that vinculin is “fully” activated by a combination of talin with other binding partners in cells.…”

Section: Resultssupporting

confidence: 84%

“…Interestingly, these mutations did not clearly reduce actin binding under mild activation conditions using VBS3(talin). Although the structure of Vt-actin is controversial [18–20], in all models, Vt binds to two actin protomers in F-actin through two surfaces. Interestingly, double substitutions at both surfaces that were based on one structural model showed a drastic reduction in actin binding [20].…”

Section: Discussionmentioning

confidence: 99%

“…Models for the Vt-actin structure have been proposed using electron microscopy [18, 19], but this structure remains controversial due to the limited resolution. A recent study using cryo-electron microscopy indicated the displacement of one of five helices in the Vt helical bundle to mediate interactions with actin [20]. In all models, Vt binds to two actin protomers in F-actin via two surfaces.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior

et al. 2017

View full text Add to dashboard Cite

The extracellular matrix (ECM) is a major regulator of cell behavior. Recent studies have indicated the importance of the physical properties of the ECM, including its stiffness, for cell migration and differentiation. Using actomyosin-generated forces, cells pull the ECM and sense stiffness via cell-ECM adhesion structures called focal adhesions (FAs). Vinculin, an actin-binding FA protein, has emerged as a major player in FA-mediated mechanotransduction. Although vinculin is important for sensing ECM stiffness, the role of vinculin binding to actin in the ECM stiffness-mediated regulation of vinculin behavior remains unknown. Here, we show that an actin binding-deficient mutation disrupts the ECM stiffness-dependent regulation of CSB (cytoskeleton stabilization buffer) resistance and the stable localization of vinculin. These results suggest that the vinculin-actin interaction participates in FA-mediated mechanotransduction.

show abstract

Section: Resultssupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior

et al. 2017

View full text Add to dashboard Cite

show abstract

“…cDNAs encoding mouse FMN2 FH2 domain, corresponding to amino acids 1137–1529 (with or without a point mutation that resulted in a I1225 to A amino acid substitution) were expressed in BL21(DE3)-Rosetta2 E. coli cells (Millipore, DarmStadt, FGR) as His6-fusion proteins, following procedures used previously (Kim et al 2015). Briefly, expression was induced in log phase cultures with 0.5 mM IPTG at 16°C.…”

Section: Methods Detailsmentioning

confidence: 99%

RETRACTED: FMN2 Makes Perinuclear Actin to Protect Nuclei during Conﬁned Migration and Promote Metastasis

Skau¹,

Fischer²,

Gurel³

et al. 2016

Cell

Self Cite

View full text Add to dashboard Cite

Summary Cell migration in confined 3D tissue microenvironments is critical for both normal physiological functions and dissemination of tumor cells. We discovered a cytoskeletal structure that prevents damage to the nucleus during migration in confined microenvironments. The formin-family actin filament nucleator FMN2 associates with and generates a perinuclear actin/focal adhesion (FA) system that is distinct from previously characterized actin/FA structures. This system controls nuclear shape and positioning in cells migrating on 2D surfaces. In confined 3D microenvironments, FMN2 promotes cell survival by limiting nuclear envelope damage and DNA double-strand breaks. We found that FMN2 is upregulated in human melanomas, and show that disruption of FMN2 in mouse melanoma cells inhibits their extravasation and metastasis to the lung. Our results indicate a critical role for FMN2 in generating a perinuclear actin/FA system that protects the nucleus and DNA from damage to promote cell survival during confined migration, and thus promote cancer metastasis.

show abstract

“…From data derived from a variety of in vivo and in vitro techniques, it was concluded that MVt "tunes" actin filament bundles by altering the architecture and flexibility (73). Using high resolution cryo-electron microscopy, a sub-nanometer three-dimensional reconstruction of the vinculin tailactin complex was obtained (74). This study suggests that upon binding to actin, the N-terminal helix H1 of Vt unfurls, leading to vinculin dimerization and actin bundling.…”

Section: Metavinculinmentioning

confidence: 99%

Mechanisms and Functions of Vinculin Interactions with Phospholipids at Cell Adhesion Sites

Izard

Brown

2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

The cytoskeletal protein vinculin is a major regulator of cell adhesion and attaches to the cell surface by binding to specific phospholipids. Structural, biochemical, and biological studies provided much insight into how vinculin binds to membranes, what components it recognizes, and how lipid binding is regulated. Here we discuss the roles and mechanisms of phospholipids in regulating the structure and function of vinculin and of its muscle-specific metavinculin splice variant. A full appreciation of these processes is necessary for understanding how vinculin regulates cell motility, migration, and wound healing, and for understanding of its role in cancer and cardiovascular diseases.Inositol phospholipids are crucial regulators of cell physiology, and their head group interactions play fundamental roles in controlling membrane/cytosol interfaces. In addition to signal transduction at the cell surface, these lipids regulate a range of cellular processes including membrane trafficking, polarity, nuclear events, the permeability and transport functions of membranes, and cytoskeletal organization. Phosphoinositides are spatially and temporally regulated at sites of actin assembly andcytoskeletonremodeling.Phosphatidylinositol4,5-bisphosphate (PIP 2 ) 2 is a precursor of the second messengers diacylglycerol, inositol 3,4,5-trisphosphate, and phosphatidyl 3,4,5-trisphosphate and is crucial in the organization of the actin cytoskeleton at the plasma membrane (1). PIP 2 participates in the recruitment and activation of a wide variety of adaptor proteins and actin regulatory proteins. During the assembly of focal adhesions (FAs), the cytoskeletal protein talin recruits an isoform of phosphatidylinositol 4-phosphate 5-kinase (PIP 5 K␥90) that catalyzes the phosphorylation of phosphatidyl 4-phosphate to generate a local enrichment of PIP 2 (2-4). This localized pool of PIP 2 is critical for the subsequent recruitment of FA proteins and maturation of the FAs (5, 6). Vinculin is an early and essential component of nascent cell-matrix adhesion and acts as a scaffold by binding to several actin-organizing proteins. In mature FAs, vinculin is a key component of the "molecular clutch" that mediates the transmission of force from cytoplasmic F-actin to membrane-bound integrins (7-9). Cytosolic vinculin mainly adopts a closed conformation that is kept inactive through extensive hydrophobic interactions of its globular 91-kDa head (VH, residues 1-840) that harbors binding sites for talin, ␣-actinin, or ␣-catenin to VH (10 -13) and its 21-kDa tail (Vt, residues 879 -1066) domains (14, 15), which are connected via a proline-rich linker (Fig. 1, A and B). The intramolecular VH-Vt interaction masks the binding sites of F-actin (16, 17), PIP 2 (18), and raver1 (19,20) to Vt. Cytoplasmic vinculin must be recruited to FAs and activated to expose ligandbinding sites through a complex process that is not completely understood (21).Vinculin is essential during development due to its role in regulating adhesion and motility, as well as cel...

show abstract

The Structural Basis of Actin Organization by Vinculin and Metavinculin

Cited by 51 publications

References 72 publications

Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior

Vinculin association with actin cytoskeleton is necessary for stiffness-dependent regulation of vinculin behavior

RETRACTED: FMN2 Makes Perinuclear Actin to Protect Nuclei during Conﬁned Migration and Promote Metastasis

Mechanisms and Functions of Vinculin Interactions with Phospholipids at Cell Adhesion Sites

Contact Info

Product

Resources

About