Bradykinin induces formation of vesicle-like structures containing vinculin and PtdIns(4,5)P<sub>2</sub>in renal papillary collecting duct cells

Márquez, Marı́a Gabriela; Fernández-Tomé, Marı́a del Carmen; Favale, Nicolás Octavio; Pescio, Lucila Gisele; Sterín-Speziale, Norma

doi:10.1152/ajprenal.00062.2009

Cited by 4 publications

(1 citation statement)

References 47 publications

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“…While association of Vcl with PIP 2 has been implicated in numerous processes including cell spreading, cell migration, FA turnover, and force transduction (Chandrasekar et al, 2005; Chinthalapudi et al, 2014; Diez et al, 2009; Saunders et al, 2006), the Vcl constructs used in these studies may not be selective in their disruption of PIP 2 -binding. Additionally, PIP 2 has been reported to regulate Vcl phosphorylation (Ziegler et al, 2002) and trafficking (Halstead et al, 2010; Marquez et al, 2009). …”

Section: Introductionmentioning

confidence: 99%

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

et al. 2017

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Summary Vinculin, a scaffolding protein that localizes to focal adhesions (FA) and adherens junctions, links the actin cytoskeleton to the adhesive super-structure. While vinculin binds to a number of cytoskeletal proteins, it can also associate with phosphatidylinositol 4,5-bisphosphate (PIP2) to drive membrane association. To generate a structural model for PIP2-dependent interaction of vinculin with the lipid bilayer, we conducted lipid-association, nuclear magnetic resonance, and computational modeling experiments. We find that two basic patches on the vinculin tail drive membrane association: the basic collar specifically recognizes PIP2, while the basic ladder drives association with the lipid bilayer. Vinculin mutants with defects in PIP2-dependent liposome association were then expressed in vinculin knockout murine embryonic fibroblasts. Results from these analyses indicate that PIP2-binding is not required for localization of vinculin to FAs or FA strengthening but is required for vinculin activation and turnover at FAs to promote its association with the force-transduction FA nanodomain.

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Section: Introductionmentioning

confidence: 99%

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

et al. 2017

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Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway

Márquez

Brandán

Guaytima

et al. 2014

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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In epithelial cells, vinculin is enriched in cell adhesion structures but is in equilibrium with a large cytosolic pool. It is accepted that when cells adhere to the extracellular matrix, a part of the soluble cytosolic pool of vinculin is recruited to specialized sites on the plasma membrane called focal adhesions (FAs) by binding to plasma membrane phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2). We have previously shown that bradykinin (BK) induces both a reversible dissipation of vinculin from FAs, by the phospholipase C (PLC)-mediated hydrolysis of PtdIns(4,5)P2, and the concomitant internalization of vinculin. Here, by using an immunomagnetic method, we isolated vinculin-containing vesicles induced by BK stimulation. By analyzing the presence of proteins involved in vesicle traffic, we suggest that vinculin can be delivered in the site of FA reassembly by a vesicular endocytic recycling pathway. We also observed the formation of vesicle-like structures containing vinculin in the cytosol of cells treated with lipid membrane-affecting agents, which caused dissipation of FAs due to their deleterious effect on membrane microdomains where FAs are inserted. However, these vesicles did not contain markers of the recycling endosomal compartment. Vinculin localization in vesicles has not been reported before, and this finding challenges the prevailing model of vinculin distribution in the cytosol. We conclude that the endocytic recycling pathway of vinculin could represent a physiological mechanism to reuse the internalized vinculin to reassembly new FAs, which occurs after long time of BK stimulation, but not after treatment with membrane-affecting agents.

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The inhibition of sphingomyelin synthase 1 activity induces collecting duct cells to lose their epithelial phenotype

Brandán

Guaytima

Favale

et al. 2018

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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Epithelial tissue requires that cells attach to each other and to the extracellular matrix by the assembly of adherens junctions (AJ) and focal adhesions (FA) respectively. We have previously shown that, in renal papillary collecting duct (CD) cells, both AJ and FA are located in sphingomyelin (SM)-enriched plasma membrane microdomains. In the present work, we investigated the involvement of SM metabolism in the preservation of the epithelial cell phenotype and tissue organization. To this end, primary cultures of renal papillary CD cells were performed. Cultured cells preserved the fully differentiated epithelial phenotype as reflected by the presence of primary cilia. Cells were then incubated for 24h with increasing concentrations of D609, a SM synthase (SMS) inhibitor. Knock-down experiments silencing SMS 1 and 2 were also performed. By combining biochemical and immunofluorescence studies, we found experimental evidences suggesting that, in CD cells, SMS 1 activity is essential for the preservation of cell-cell adhesion structures and therefore for the maintenance of CD tissue/tubular organization. The inhibition of SMS 1 activity induced CD cells to lose their epithelial phenotype and to undergo an epithelial-mesenchymal transition (EMT) process.

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Bradykinin induces formation of vesicle-like structures containing vinculin and PtdIns(4,5)P₂in renal papillary collecting duct cells

Cited by 4 publications

References 47 publications

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway

The inhibition of sphingomyelin synthase 1 activity induces collecting duct cells to lose their epithelial phenotype

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Bradykinin induces formation of vesicle-like structures containing vinculin and PtdIns(4,5)P2in renal papillary collecting duct cells

Cited by 4 publications

References 47 publications

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization

Physiologically induced restructuring of focal adhesions causes mobilization of vinculin by a vesicular endocytic recycling pathway

The inhibition of sphingomyelin synthase 1 activity induces collecting duct cells to lose their epithelial phenotype

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Bradykinin induces formation of vesicle-like structures containing vinculin and PtdIns(4,5)P₂in renal papillary collecting duct cells