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.
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24 August 1998
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August 24 1998
The Ras Target AF-6 is a Substrate of the Fam Deubiquitinating Enzyme
Shinichiro Taya,
Shinichiro Taya
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Takaharu Yamamoto,
Takaharu Yamamoto
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Kyoko Kano,
Kyoko Kano
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Yoji Kawano,
Yoji Kawano
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Akihiro Iwamatsu,
Akihiro Iwamatsu
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Tomoko Tsuchiya,
Tomoko Tsuchiya
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Keiji Tanaka,
Keiji Tanaka
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Masami Kanai-Azuma,
Masami Kanai-Azuma
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Stephen A. Wood,
Stephen A. Wood
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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John S. Mattick,
John S. Mattick
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Kozo Kaibuchi
Kozo Kaibuchi
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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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Shinichiro Taya
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Takaharu Yamamoto
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Kyoko Kano
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Yoji Kawano
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Akihiro Iwamatsu
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Tomoko Tsuchiya
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Keiji Tanaka
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Masami Kanai-Azuma
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Stephen A. Wood
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
John S. Mattick
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Kozo Kaibuchi
*Division of Signal Transduction, Nara Institute of Science and Technology, Ikoma 630-0101, Japan; ‡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
Address all correspondence and proofs to Kozo Kaibuchi, M.D. & Ph.D., Division of Signal Transduction, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan. Tel.: 81-743-72-5440. Fax: 81-743-72-5449. E-mail: [email protected]
Received:
May 28 1998
Revision Received:
July 09 1998
Online ISSN: 1540-8140
Print ISSN: 0021-9525
1998
J Cell Biol (1998) 142 (4): 1053–1062.
Article history
Received:
May 28 1998
Revision Received:
July 09 1998
Citation
Shinichiro Taya, Takaharu Yamamoto, Kyoko Kano, Yoji Kawano, Akihiro Iwamatsu, Tomoko Tsuchiya, Keiji Tanaka, Masami Kanai-Azuma, Stephen A. Wood, John S. Mattick, Kozo Kaibuchi; The Ras Target AF-6 is a Substrate of the Fam Deubiquitinating Enzyme . J Cell Biol 24 August 1998; 142 (4): 1053–1062. doi: https://doi.org/10.1083/jcb.142.4.1053
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