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Domain analysis of supervillin, an F-actin bundling plasma membrane protein with functional nuclear localization signals.

Journal of cell science | Jul 2, 1999

A growing number of actin-associated membrane proteins have been implicated in motile processes, adhesive interactions, and signal transduction to the cell nucleus. We report here that supervillin, an F-actin binding protein originally isolated from bovine neutrophil plasma membranes, contains functional nuclear targeting signals and localizes at or near vinculin-containing focal adhesion plaques in COS7-2 and CV1 cells. Overexpression of full-length supervillin in these cells disrupts the integrity of focal adhesion plaques and results in increased levels of F-actin and vinculin. Localization studies of chimeric proteins containing supervillin sequences fused with the enhanced green fluorescent protein indicate that: (1) the amino terminus promotes F-actin binding, targeting to focal adhesions, and limited nuclear localization; (2) the dominant nuclear targeting signal is in the center of the protein; and (3) the carboxy-terminal villin/gelsolin homology domain of supervillin does not, by itself, bind tightly to the actin cytoskeleton in vivo. Overexpression of chimeras containing both the amino-terminal F-actin binding site(s) and the dominant nuclear targeting signal results in the formation of large nuclear bundles containing F-actin, supervillin, and lamin. These results suggest that supervillin may contribute to cytoarchitecture in the nucleus, as well as at the plasma membrane.

Pubmed ID: 10362542 RIS Download

Mesh terms: Actins | Animals | Base Sequence | Binding Sites | COS Cells | Cattle | Cell Adhesion | Cell Line | Cytoskeleton | DNA Primers | Gene Expression | Green Fluorescent Proteins | Lamins | Luminescent Proteins | Membrane Proteins | Microfilament Proteins | Nuclear Localization Signals | Nuclear Proteins | Phenotype | Recombinant Fusion Proteins | Vinculin

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Associated grants

  • Agency: NIGMS NIH HHS, Id: R01 GM033048
  • Agency: NICHD NIH HHS, Id: 5T32HD07312-12
  • Agency: NIGMS NIH HHS, Id: GM33048

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