Wulfkuhle, J. D.Donina, I. E.Stark, N. H.Pope, Robert K.Pestonjamasp, Kersi N.Niswonger, M. L.Luna, Elizabeth J.2022-08-232022-08-231999-06-112007-11-28J Cell Sci. 1999 Jul;112 ( Pt 13):2125-36. <a href="http://jcs.biologists.org/cgi/reprint/112/13/2125">Link to article on publisher's website</a>0021-9533 (Print)10362542https://hdl.handle.net/20.500.14038/50775A 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.en-USActinsAnimalsBase SequenceBinding SitesCOS CellsCattleCell AdhesionCell LineCytoskeletonDNA PrimersGene ExpressionGreen Fluorescent ProteinsLaminsLuminescent ProteinsMembrane ProteinsMicrofilament ProteinsNuclear Localization SignalsNuclear ProteinsPhenotypeRecombinant Fusion ProteinsVinculinCell BiologyLife SciencesMedicine and Health SciencesDomain analysis of supervillin, an F-actin bundling plasma membrane protein with functional nuclear localization signalsJournal Articlehttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1299&context=wfc_pp&unstamped=1https://escholarship.umassmed.edu/wfc_pp/300397390wfc_pp/300