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

Peter M. Thompson, Srinivas Ramachandran, Lindsay B. Case, Caitlin E. Tolbert, Arpit Tandon, Mihir Pershad, Nikolay Dokholyan, Clare M. Waterman, Sharon L. Campbell

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Vinculin, a scaffolding protein that localizes to focal adhesions (FAs) 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.

Original languageEnglish (US)
Pages (from-to)264-275
Number of pages12
JournalStructure
Volume25
Issue number2
DOIs
StatePublished - Feb 7 2017

Fingerprint

Vinculin
Structural Models
Focal Adhesions
Membranes
Lipid Bilayers
Adherens Junctions
Cytoskeletal Proteins
Phosphatidylinositols
Actin Cytoskeleton
Liposomes
Adhesives
Magnetic Resonance Spectroscopy
Fibroblasts
Lipids

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Thompson, P. M., Ramachandran, S., Case, L. B., Tolbert, C. E., Tandon, A., Pershad, M., ... Campbell, S. L. (2017). A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization. Structure, 25(2), 264-275. https://doi.org/10.1016/j.str.2016.12.002
Thompson, Peter M. ; Ramachandran, Srinivas ; Case, Lindsay B. ; Tolbert, Caitlin E. ; Tandon, Arpit ; Pershad, Mihir ; Dokholyan, Nikolay ; Waterman, Clare M. ; Campbell, Sharon L. / A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization. In: Structure. 2017 ; Vol. 25, No. 2. pp. 264-275.
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abstract = "Vinculin, a scaffolding protein that localizes to focal adhesions (FAs) 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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Thompson, PM, Ramachandran, S, Case, LB, Tolbert, CE, Tandon, A, Pershad, M, Dokholyan, N, Waterman, CM & Campbell, SL 2017, 'A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization', Structure, vol. 25, no. 2, pp. 264-275. https://doi.org/10.1016/j.str.2016.12.002

A Structural Model for Vinculin Insertion into PIP2-Containing Membranes and the Effect of Insertion on Vinculin Activation and Localization. / Thompson, Peter M.; Ramachandran, Srinivas; Case, Lindsay B.; Tolbert, Caitlin E.; Tandon, Arpit; Pershad, Mihir; Dokholyan, Nikolay; Waterman, Clare M.; Campbell, Sharon L.

In: Structure, Vol. 25, No. 2, 07.02.2017, p. 264-275.

Research output: Contribution to journalArticle

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AU - Thompson, Peter M.

AU - Ramachandran, Srinivas

AU - Case, Lindsay B.

AU - Tolbert, Caitlin E.

AU - Tandon, Arpit

AU - Pershad, Mihir

AU - Dokholyan, Nikolay

AU - Waterman, Clare M.

AU - Campbell, Sharon L.

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