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Biochemical and Structural Analysis of Kinetochore Histone-Fold Complexes

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The Mitotic Spindle

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1413))

Abstract

The kinetochore connects chromosomes to microtubules during mitosis and therefore plays an essential role in faithful chromosome segregation. It is built at the centromeric region of the chromosome and is comprised of many protein complexes. CENP-S, -T, -W, and -X are kinetochore components with histone-folds. These proteins play important roles in establishment of kinetochore chromatin. Similar to canonical histones, these kinetochore histone-fold proteins form heteromeric complexes (CENP-S/CENP-X complex and CENP-T/CENP-W complex) and bind DNA in sequence independent manner. In addition, they form a CENP-T-W-S-X heterotetrameric complex and bind DNA in a manner that is different from both CENP-S-X and CENP-T-W. To understand how kinetochores form and function it is necessary to characterize the components in detail. Here, we describe our approaches in purification and characterization of the kinetochore histone-fold complexes.

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Acknowledgements

We thank members of the Fukagawa lab in particular Kozo Takeuchi for his help with sample preparation and characterization. We also acknowledge Kiyomi Kita and Mayumi Takahashi for their contributions.

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Authors and Affiliations

  1. Department of Molecular Genetics, National Institute of Genetics and Graduate University for Advanced Studies (SOKENDAI), 1111 Yata, Mishima, Shizuoka, 411-8540, Japan

    Tatsuya Nishino & Tatsuo Fukagawa

  2. Department of Biological Science and Technology, Graduate School of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, 125-8585, Japan

    Tatsuya Nishino

  3. Graduate School of Frontier Biosciences, Osaka University, Suita 565-0871, Japan

    Tatsuo Fukagawa

Authors
  1. Tatsuya Nishino
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  2. Tatsuo Fukagawa
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Correspondence to Tatsuo Fukagawa .

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Editors and Affiliations

  1. Massachusetts Institute of Technolo, Cambridge, Massachusetts, USA

    Paul Chang

  2. Dept of Cell & Developmental Biolog, Vanderbilt University, Nashville, Tennessee, USA

    Ryoma Ohi

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Nishino, T., Fukagawa, T. (2016). Biochemical and Structural Analysis of Kinetochore Histone-Fold Complexes. In: Chang, P., Ohi, R. (eds) The Mitotic Spindle. Methods in Molecular Biology, vol 1413. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3542-0_9

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  • DOI: https://doi.org/10.1007/978-1-4939-3542-0_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3540-6

  • Online ISBN: 978-1-4939-3542-0

  • eBook Packages: Springer Protocols

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