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Inn1 couples contraction of the actomyosin ring to membrane ingression during cytokinesis in budding yeast

Nature Cell Biology volume 10, pages 395–406 (2008)Cite this article

Abstract

By rapidly depleting each of the essential budding yeast proteins of unknown function, we identified a novel factor that we call Inn1, which associates with the contractile actomyosin ring at the end of mitosis and is needed for cytokinesis. We show that Inn1 has a C2 domain at the amino terminus of the protein that is required for ingression of the plasma membrane, whereas the remainder of the protein recruits Inn1 to the actomyosin ring. The lethal effects of deleting the INN1 gene can be suppressed by artificial fusion of the C2 domain to other components of the actomyosin ring, restoring membrane ingression on contraction of the actomyosin ring. Our data indicate that recruitment of the C2 domain of Inn1 to the contractile actomyosin ring is crucial for ingression of the plasma membrane during cytokinesis in budding yeast.

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Figure 1: Inn1 is required for cell division and associates with the contractile actomyosin ring.
Figure 2: Localization of Inn1 is dependent on components of the actomyosin ring.
Figure 3: Inn1 is not required for assembly or contraction of the actomyosin ring, or for correct localization of the septin component Cdc12.
Figure 4: Inn1 is required for division of the cytoplasm at the end of mitosis.
Figure 5: Inn1 is required for ingression of the plasma membrane during cytokinesis.
Figure 6: The Inn1 protein has a C2 domain at its N-terminus, and the rest of the protein is rich in PXXP motifs.
Figure 7: The C2 domain of Inn1 is essential for ingression of the plasma membrane during cytokinesis and the remainder of the protein is required to recruit Inn1 to the bud-neck.
Figure 8: Artificial recruitment of the C2 domain of Inn1 to the actomyosin ring is sufficient to allow ingression of the plasma membrane in cells lacking the INN1 gene.

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Acknowledgements

This work was funded by Cancer Research U.K., from whom K.L. received a Senior Cancer Research Fellowship, and by the EMBO Young Investigator Programme. A.S.D. received a Marie Curie personal training fellowship from the European Union. We are very grateful to S. Bagley for much advice and help with time-lapse video microscopy, B. Glick for advice regarding the use of concanavalin A, J. Wiedenmann for the eQFP construct, and E. Schiebel for plasmids and helpful comments on the manuscript. A.S.D. thanks A. Sanchez-Sanfructuoso and J. Diaz-Marcos for their support.

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Author notes

  1. Richard Jones

    Present address: Current address: NextGen Sciences, Inc., 4401 Varsity Drive, Suite E, Ann Arbor, MI 48108, USA.,

  2. Gislene Pereira

    Present address: Current address: German Cancer Research Centre, Im Neuenheimer Feld 581, 69120 Heidelberg, Germany.,

  3. Ricky Edmondson

    Present address: Current address: Myeloma Institute for Research and Therapy, University of Arkansas for Medical Sciences, 4301 W.Markham #776, Little Rock, AR 72205, USA.,

Authors and Affiliations

  1. Cancer Research U.K., Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK

    Alberto Sanchez-Diaz, Vanessa Marchesi, Stephen Murray, Gislene Pereira, Terry Allen & Karim Labib

  2. FDA-NCTR, 3900 NCTR Road, Jefferson, 72079, AR, USA

    Richard Jones & Ricky Edmondson

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Contributions

V. M. generated the strains that are described in Supplementary Information, Fig. S1a and b, and screened these strains for defects in cell cycle progression to generate the corrsponding data in Supplementary Information, Fig. 1c. S. M. performed the electron microscopy that is summarized in Fig. 4a. R. J. was responsible for the mass spectrometry data in Fig. 1e. G. P. made the initial observations of the localization of Inn–GFP and taught A. S. D. how to prepare samples for fluorescence microscopy. K. L. made the strains shown in Fig. 8a, b, c. A. S. D. performed all other experiments.

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Correspondence to Karim Labib.

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Sanchez-Diaz, A., Marchesi, V., Murray, S. et al. Inn1 couples contraction of the actomyosin ring to membrane ingression during cytokinesis in budding yeast. Nat Cell Biol 10, 395–406 (2008). https://doi.org/10.1038/ncb1701

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