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

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

Authors

  • Wulfkuhle JD
  • Donina IE
  • Stark NH
  • Pope RK
  • Pestonjamasp KN
  • Niswonger ML
  • Luna EJ

Journal

Journal of cell science

Publication Data

July 2, 1999

Associated Grants

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

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