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Molecular and Cellular Biochemistry

, Volume 320, Issue 1–2, pp 53–66 | Cite as

Truncated form of tenascin-X, XB-S, interacts with mitotic motor kinesin Eg5

  • Toshiya Endo
  • Hiroyoshi Ariga
  • Ken-ichi Matsumoto
Article

Abstract

XB-S is a protein with an amino-terminal-truncated form of tenascin-X (TNXB). However, the precise roles of XB-S in vivo are unknown. In this study, to determine the role of XB-S in vivo, we screened XB-S-binding proteins. FLAG-tagged XB-S was transiently introduced into 293T cells. Then its associated proteins were purified by immunoprecipitation using an anti-FLAG antibody and its components were identified by mass spectrometric analyses. Mitotic motor kinesin Eg5 was identified in the immunoprecipitates. XB-S and Eg5 proteins were co-localized in the cytoplasm in interphase and mitosis, but XB-S did not localize on mitotic spindle microtubules, on which Eg5 prominently localized in mitosis. As for Eg5 binding to XB-S, glutathione S-transferase-fused XB-S expressed in vitro directly bound to full-length Eg5 translated in reticulocyte lysate, and the XB-S-binding region was located in the motor domain of Eg5. Furthermore, during cell cycle progression XB-S showed a similar expression profile to that of Eg5. These results suggest possible involvement of XB-S in the function of Eg5.

Keywords

Mitosis Spindle Tenascin-X TNXA TNXB XB-S 

Notes

Acknowledgments

We thank Kiyomi Takaya for her technical assistance. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Toshiya Endo
    • 1
  • Hiroyoshi Ariga
    • 2
  • Ken-ichi Matsumoto
    • 3
  1. 1.Graduate School of Life ScienceHokkaido UniversitySapporoJapan
  2. 2.Department of Molecular Biology, Graduate School of Pharmaceutical SciencesHokkaido UniversitySapporoJapan
  3. 3.Laboratory of Molecular Biology, Faculty of Advanced Life ScienceHokkaido UniversitySapporoJapan

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