FEBS Letters
Volume 582, Issue 8 , Pages 1189-1196, 9 April 2008

Affixin activates Rac1 via βPIX in C2C12 myoblast

Edited by Berend Wieringa

  • Chie Matsuda

      Affiliations

    • Neuroscience Research Institute, AIST, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan
    • Department of Neuromuscular Research, National Institute of Neuroscience, NCNP, Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan
    • Corresponding Author InformationCorresponding author. Fax: +81 29 861 6482.
    • ,
  • Kimihiko Kameyama

      Affiliations

    • Neuroscience Research Institute, AIST, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan
    • ,
  • Atsushi Suzuki

      Affiliations

    • Department of Molecular Biology, Yokohama City University, Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan
    • ,
  • Wataru Mishima

      Affiliations

    • Department of Internal Medicine and Clinical Immunology, Yokohama City University, Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan
    • ,
  • Satoshi Yamaji

      Affiliations

    • Department of Internal Medicine and Clinical Immunology, Yokohama City University, Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan
    • ,
  • Harumasa Okamoto

      Affiliations

    • Neuroscience Research Institute, AIST, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan
    • ,
  • Ichizo Nishino

      Affiliations

    • Department of Neuromuscular Research, National Institute of Neuroscience, NCNP, Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan
    • ,
  • Yukiko K. Hayashi

      Affiliations

    • Department of Neuromuscular Research, National Institute of Neuroscience, NCNP, Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan

Received 25 December 2007; accepted 31 January 2008. published online 04 March 2008.

Abstract 

Affixin/β-parvin is an integrin-linked kinase (ILK)-binding focal adhesion protein highly expressed in skeletal muscle and heart. To elucidate the possible role of affixin in skeletal muscle, we established stable C2C12 cell line expressing T7-tagged human affixin (C2C12-affixin cells). Exogenous expression of affixin promotes lamellipodium formation where affixin, ILK αp21-activated kinase (PAK)-interactive exchange factor (PIX) and βPIX accumulate. The association of affixin and βPIX was confirmed by immunoprecipitation and pull down assay. In C2C12-affixin cells, an increased level of activated Rac1 but not Cdc42 was observed, and mutant βPIX lacking guanine nucleotide exchange factor activity inhibited lamellipodium formation. These results suggest that affixin is involved in reorganization of subsarcolemmal cytoskeletal actin by activation of Rac1 through α and βPIXs in skeletal muscle.

Structured summary


MINT-6179203, MINT-6179212, MINT-6178859, MINT-6178812, MINT-6178832, MINT-6178843:Affixin (uniprotkb:Q9HBI1) physically interacts (MI:0218) with βpix (uniprotkb:Q9ES28) by coimmunoprecipitation (MI:0019)

MINT-6179221:Affixin (uniprotkb:Q9HBI1) physically interacts (MI:0218) with αpix (uniprotkb:Q8K4I3) by coimmunoprecipitation (MI:0019)

MINT-6178962, MINT-6178983:Affixin (uniprotkb:Q9HBI1) physically interacts (MI:0218) with βpix (uniprotkb:Q9ES28) by pull-down (MI:0096)

MINT-6179002, MINT-6179021:Affixin (uniprotkb:Q9HBI1) binds (MI:0407) βpix (uniprotkb:Q9ES28) by pull-down (MI:0096)

MINT-6179039:PAK1 (uniprotkb:Q13153) physically interacts (MI:0218) with Rac1 (uniprotkb:P63001) by pull-down (MI:0096)

MINT-6179054:PAK1 (uniprotkb:Q13153) physically interacts (MI:0218) with Cdc42 (uniprotkb:P70766) by pull-down (MI:0096)

MINT-6178790:Affixin (uniprotkb:Q9HBI1) and αpix (uniprotkb:Q8K4I3) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

MINT-6178760:Affixin (uniprotkb:Q9HBI1) and βpix (uniprotkb:Q9ES28) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

MINT-6178801:Affixin (uniprotkb:Q9HBI1) and dysferlin (uniprotkb:Q9ESD7) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

MINT-6178779:Affixin (uniprotkb:Q9HBI1) and ILK (uniprotkb:O55222) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

Abbreviations: ILK, integrin-linked kinase, PIX, PAK-interactive exchange factor, C2C12-affixin, stable C2C12 cell line expressing T7-tagged human affixin, GEF, guanine nucleotide exchange factor, GST, gulutathione S-transferase, CH, caponin-homology, DH, Dbl-homology, PH, pleckstrin-homology, PAK, p21-activated kinase, CHO, Chinese hamster ovary

Keywords: Affixin/β-parvin, Lamellipodia, βPIX, Cytoskeletal actin

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PII: S0014-5793(08)00168-3

doi:10.1016/j.febslet.2008.01.064

FEBS Letters
Volume 582, Issue 8 , Pages 1189-1196, 9 April 2008

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