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

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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.

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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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Authors and Affiliations

  1. Graduate School of Life Science, Hokkaido University, Kita-ku, Sapporo, 060-0812, Japan

    Toshiya Endo

  2. Department of Molecular Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo, 060-0812, Japan

    Hiroyoshi Ariga

  3. Laboratory of Molecular Biology, Faculty of Advanced Life Science, Hokkaido University, Kita 12, Nishi 6, Kita-ku, Sapporo, 060-0812, Japan

    Ken-ichi Matsumoto

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Correspondence to Ken-ichi Matsumoto.

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Endo, T., Ariga, H. & Matsumoto, Ki. Truncated form of tenascin-X, XB-S, interacts with mitotic motor kinesin Eg5. Mol Cell Biochem 320, 53–66 (2009). https://doi.org/10.1007/s11010-008-9898-y

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