© The Rockefeller University Press,
0021-9525/1999//59 $5.00
The Journal of Cell Biology, Volume 144, Number 1,
, 1999 59-69
Vinexin: A Novel Vinculin-binding Protein with Multiple SH3 Domains Enhances Actin Cytoskeletal Organization
Noriyuki Kioka*,
,
Shohei Sakata
,
Takeshi Kawauchi
,
Teruo Amachi
,
Steven K. Akiyama*,
,
Kenji Okazaki||,
Christopher Yaen*,
Kenneth M. Yamada*, and
Shin-ichi Aota*,||
* Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892;
Laboratory of Biochemistry, Division of Applied Life Science, Kyoto University, Kyoto 606-01, Japan;
Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Science, Research Triangle Park, North Carolina 27709; and || Biomolecular Engineering Research Institute, Osaka 565-0874, Japan
Using the yeast two-hybrid system and an in vitro binding assay, we have identified a novel protein termed vinexin as a vinculin-binding protein. By Northern blotting, we identified two types of vinexin mRNA that were 3 and 2 kb in length. Screening for full-length cDNA clones and sequencing indicated that the two mRNA encode 82- and 37-kD polypeptides termed vinexin
and β, respectively. Both forms of vinexin share a common carboxyl-terminal sequence containing three SH3 domains. The larger vinexin
contains an additional amino-terminal sequence. The interaction between vinexin and vinculin was mediated by two SH3 domains of vinexin and the proline-rich region of vinculin. When expressed, vinexin
and β localized to focal adhesions in NIH 3T3 fibroblasts, and to cell–cell junctions in epithelial LLC-PK1 cells. Furthermore, expression of vinexin increased focal adhesion size. Vinexin
also promoted upregulation of actin stress fiber formation. In addition, cell lines stably expressing vinexin β showed enhanced cell spreading on fibronectin. These data identify vinexin as a novel focal adhesion and cell– cell adhesion protein that binds via SH3 domains to the hinge region of vinculin, which can enhance actin cytoskeletal organization and cell spreading.
Key Words: actin cytoskeleton cell adhesion focal adhesion SH3 domain vinculin
Abbreviations used in this paper: GFP, green fluorescent protein; GST, glutathione S-transferase; VASP, vasodilator-stimulated phosphoprotein.
Address correspondence to Shin-ichi Aota, Biomolecular Engineering Research Institute, Suita, 6-2-3 Furuedai, Osaka 565-0874, Japan. Tel.: 81-6-872-8208. Fax: 81-6-872-8219. E-mail: aota{at}beri.co.jp

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