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J. Cell Biol.,
Volume 142, Number 4, August 24, 1998 1053-1062
* Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; The Ras target AF-6 has been shown to
serve as one of the peripheral components of cell-cell
adhesions, and is thought to participate in cell-cell adhesion regulation downstream of Ras. We here purified
an AF-6-interacting protein with a molecular mass of
~220 kD (p220) to investigate the function of AF-6 at cell-cell adhesions. The peptide sequences of p220
were identical to the amino acid sequences of mouse
Fam. Fam is homologous to a deubiquitinating enzyme
in Drosophila, the product of the fat facets gene. Recent
genetic analyses indicate that the deubiquitinating activity of the fat facets product plays a critical role in controlling the cell fate. We found that Fam accumulated
at the cell-cell contact sites of MDCKII cells, but not at
free ends of plasma membranes. Fam was partially
colocalized with AF-6 and interacted with AF-6 in vivo
and in vitro. We also showed that AF-6 was ubiquitinated in intact cells, and that Fam prevented the ubiquitination of AF-6.
Central Laboratories for
Key Technology, Kirin Brewery Company Limited, Kanazawa-ku, Yokohama 236-0004, Japan; § Metropolitan Institute of
Medical Science, Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Bunkyo-ku,
Tokyo 113-0021, Japan; Centre for Molecular and Cellular Biology, University of Queensland, St. Lucia, Queensland 4072, Australia; and ¶ Department of Biochemistry, University of Adelaide, Adelaide 5005, Australia
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