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The chromosomal association of condensin II is regulated by a noncatalytic function of PP2A

Nature Structural & Molecular Biology volume 16, pages 1302–1308 (2009)Cite this article

Abstract

Mitotic chromosomal assembly in vertebrates is regulated by condensin I and condensin II, which work cooperatively but have different chromosomal localization profiles and make distinct mechanistic contributions to this process. We show here that protein phosphatase 2A (PP2A), which interacts with condensin II but not condensin I, plays an essential role in targeting condensin II to chromosomes. Unexpectedly, our data indicate that PP2A acts as a recruiter protein rather than a catalytic enzyme to target condensin II to chromosomes. This recruiting activity of PP2A was inhibited by okadaic acid, but not by fostriecin, even though both molecules strongly inhibited the catalytic activity of PP2A. Additionally, we found that the chromokinesin KIF4a is also targeted to chromosomes via the noncatalytic activity of PP2A. Thus, our studies reveal a previously unknown contribution of PP2A to chromosome assembly.

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Figure 1: Condensin II dissociates from assembled chromosomes following treatment with PP2A inhibitors.
Figure 2: PP2A is required for the chromosomal localization of condensin II.
Figure 3: PP2A specifically associates with condensin II and mitotic chromosomes.
Figure 4: The phosphatase activity of PP2A is not essential for the chromosomal association of condensin II.
Figure 5: PP2A-mediated chromosomal localization of condensin II is conserved in human cells.
Figure 6: KIF4a is also localized to chromosomes via noncatalytic action of PP2A.

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Acknowledgements

We thank M. Iwabuchi and K. Ohsumi (Nagoya University) for a plasmid that expresses a nondegradable version of Xenopus Cyclin B, W. Earnshaw (University of Edinburgh) for anti-CREST, T. Hirota (Japanese Foundation for Cancer Research) for anti–condensin II, and M. Shimura for technical advice. This work was supported by a MEXT grant-in-aid from the Solution Oriented Research for Science and Technology (SORST) from the Japan Science and Technology Agency; the 21st Century Center of Excellence Program; a nuclear system to decipher operation code (DECODE); the Naito Foundation; a grant on Advanced Medical Technology from the Ministry of Health, Labour and Welfare; and the RIKEN Bioarchitect Project. A. Takemoto is a Japan Society for the Promotion of Science (JSPS) Research Fellow.

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Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba Science City, Ibaraki, Japan

    Ai Takemoto, Kazumitsu Yamaguchi, Akiko Murayama, Junn Yanagisawa & Keiji Kimura

  2. Cellular Physiology Laboratory, RIKEN and SORST, Japan Science and Technology Agency, Wako, Saitama, Japan

    Ai Takemoto & Fumio Hanaoka

  3. Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Ai Takemoto & Shigeyuki Yokoyama

  4. Cellular Dynamics Laboratory, RIKEN, Wako, Saitama, Japan

    Kazuhiro Maeshima & Naoko Imamoto

  5. Laboratory for Biological Macromolecules, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka, Japan

    Kazuhiro Maeshima

  6. TARA Center, University of Tsukuba, Tsukuba Science City, Ibaraki, Japan

    Tsuyoshi Ikehara, Akiko Murayama & Junn Yanagisawa

  7. Tropical Technology Center Ltd., Uruma, Okinawa, Japan

    Shihoko Imamura

  8. RIKEN Genomic Sciences Center, Tsurumi, Yokohama, Japan

    Shigeyuki Yokoyama

  9. Chromosome Dynamics Laboratory, RIKEN, Wako, Saitama, Japan

    Tatsuya Hirano

  10. Laboratory of Chromosome Dynamics, Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Yoshinori Watanabe

  11. Faculty of Science, Gakushuin University, Toshima-ku, Tokyo, Japan

    Fumio Hanaoka

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Contributions

A.T., K.M. and K.K. designed the study; A.M., N.I., S.Y., T.H., Y.W., F.H. and J.Y. advised; A.T. carried out most of the experiments; K.M. performed immunostaining experiments; K.Y. and K.K. performed experiments for revision; T.I. and S.I. purified PP2A; K.M. and K.K. wrote the manuscript.

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Correspondence to Keiji Kimura.

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Takemoto, A., Maeshima, K., Ikehara, T. et al. The chromosomal association of condensin II is regulated by a noncatalytic function of PP2A. Nat Struct Mol Biol 16, 1302–1308 (2009). https://doi.org/10.1038/nsmb.1708

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