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Calcineurin ensures a link between the DNA replication checkpoint and microtubule-dependent polarized growth

Nature Cell Biology volume 13, pages 234–242 (2011)Cite this article

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Abstract

Microtubules are central to eukaryotic cell morphogenesis. Microtubule plus-end tracking proteins (+TIPs) transport polarity factors to the cell cortex, thereby playing a key role in both microtubule dynamics and cell polarity. However, the signalling pathway linking +TIPs to cell polarity control remains elusive. Here we show that the fission yeast checkpoint kinase Cds1 (Chk2 homologue) delays the transition of growth polarity from monopolar to bipolar (termed NETO; new-end take-off). The +TIPs CLIP170 homologue Tip1 and kinesin Tea2 are responsible for this delay, which is accompanied by a reduction in microtubule dynamics at the cell tip. Remarkably, microtubule stabilization occurs asymmetrically, prominently at the non-growing cell end, which induces abnormal accumulation of the polarity factor Tea1. Importantly, NETO delay requires activation of calcineurin, which is carried out by Cds1, resulting in Tip1 dephosphorylation. Thus, our study establishes a critical link between calcineurin and checkpoint-dependent cell morphogenesis.

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Figure 1: Cds1 induces +TIPs-dependent NETO delay.
Figure 2: Calcineurin (CN) induces +TIPs-dependent NETO delay.
Figure 3: Cds1 and calcineurin physically interact with +TIPs.
Figure 4: Co-localization of Cds1, calcineurin and +TIPs.
Figure 5: Protein interactions with calcineurin are required for NETO delay.
Figure 6: Phosphorylation of calcineurin by Cds1 and dephosphorylation of Tip1 by calcineurin.
Figure 7: Cds1 regulates microtubule-dependent growth polarity through calcineurin and +TIPs.

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Acknowledgements

We thank D. Brunner, K. Kitamura, H. Murakami and P. Russell for strains and plasmids; K. Gull for antibody; E. Kinoshita for Phos-tag; Astellas Pharma for providing FK506; and all members of our laboratory for their help. We also thank D. Brunner, S. Martin, D. McCollum, K. Sawin, M. Yanagida and J. Hirata for discussions; and H. Iefuji, K. Ono, T. Miyakawa and K. Hirata for their support. This work was supported by grants from the Ministry of Education, Science, and Culture of Japan (to K.K. and D.H.), and by Cancer Research UK (to T.T.)

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Authors and Affiliations

  1. Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima 739-8530, Japan

    Kazunori Kume, Takayuki Koyano & Dai Hirata

  2. National Research Institute of Brewing, Higashi-Hiroshima 739-0046, Japan

    Muneyoshi Kanai

  3. Laboratory of Cell Regulation, Cancer Research-UK, London Research Institute, PO Box 123, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK

    Takashi Toda

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Experimental design and interpretation of data were conducted by all authors. K.K., T.K. and M.K. carried out experiments. K.K. and D.H. planned the project and D.H. wrote the paper with input from the co-authors. T.T. reviewed the manuscript.

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Correspondence to Dai Hirata.

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Kume, K., Koyano, T., Kanai, M. et al. Calcineurin ensures a link between the DNA replication checkpoint and microtubule-dependent polarized growth. Nat Cell Biol 13, 234–242 (2011). https://doi.org/10.1038/ncb2166

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