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Drosophila katanin is a microtubule depolymerase that regulates cortical-microtubule plus-end interactions and cell migration

Nature Cell Biology volume 13, pages 361–369 (2011)Cite this article

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Abstract

Regulation of microtubule dynamics at the cell cortex is important for cell motility, morphogenesis and division. Here we show that the Drosophila katanin Dm-Kat60 functions to generate a dynamic cortical-microtubule interface in interphase cells. Dm-Kat60 concentrates at the cell cortex of S2 Drosophila cells during interphase, where it suppresses the polymerization of microtubule plus-ends, thereby preventing the formation of aberrantly dense cortical arrays. Dm-Kat60 also localizes at the leading edge of migratory D17 Drosophila cells and negatively regulates multiple parameters of their motility. Finally, in vitro, Dm-Kat60 severs and depolymerizes microtubules from their ends. On the basis of these data, we propose that Dm-Kat60 removes tubulin from microtubule lattice or microtubule ends that contact specific cortical sites to prevent stable and/or lateral attachments. The asymmetric distribution of such an activity could help generate regional variations in microtubule behaviours involved in cell migration.

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Figure 1: Dm-Kat60 targets the cell cortex of interphase cells.
Figure 2: Depletion of Dm-Kat60 causes significant microtubule curling and bundling beneath the cortex.
Figure 3: Automated tracking and quantitative analysis of microtubule plus-end organization and dynamics in control and Dm-Kat60 RNAi-treated cells.
Figure 4: Dm-Kat60 targets to the leading edge of motile D17 cells and negatively regulates their migration.
Figure 5: Dm-Kat60 negatively regulates multiple parameters of D17-cell motility.
Figure 6: Dm-Kat60 negatively regulates actin protrusions at the cell edge.
Figure 7: Dm-Kat60 severs and depolymerizes microtubules from their ends.

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Acknowledgements

D.J.S., U.R. and D.Z. were supported by NIH grant no. R01-GM065940 (to D.J.S.). S.L.R., K.D.G. and J.D.C. were supported by March of Dimes grant no. 1-FY08-429 and NIH grant no. R01 GM081645 (both to S.L.R.). S.F.S. and A.M. were supported by NIH grant no. R01-GM086536 (to A.M.). J.L.R. and J.D.D-V. were supported by March of Dimes grant no. 5-FY09-47 (to J.L.R.). H.J.S., A.B.A., E.L. and T.R. were supported by NIH grant no. R01-GM083338 (to H.J.S.).

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Author notes

  1. Kyle D. Grode, Shannon F. Stewman, Juan Daniel Diaz-Valencia, Jennifer L. Ross, Ao Ma and Stephen L. Rogers: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Physiology and Biophysics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA

    Dong Zhang, Shannon F. Stewman, Emily Liebling, Uttama Rath, Tania Riera, Ana B. Asenjo, Hernando J. Sosa, Ao Ma & David J. Sharp

  2. Department of Biology and the Center for Carolina Genome Sciences, University of North Carolina, Chapel Hill, 421 Fordham Hall, CB#3280, Chapel Hill, North Carolina 27599, USA

    Kyle D. Grode, Joshua D. Currie & Stephen L. Rogers

  3. Department of Physics, University or Massachusetts, Amherst, 302 Hasbrouck Laboratory, Amherst, Massachusetts 01003, USA

    Juan Daniel Diaz-Valencia & Jennifer L. Ross

  4. Department of Cell Biology and Anatomy and the Arizona Cancer Center, University of Arizona, Tuscon, 1515 N. Campbell Avenue, Tuscon, Arizona 85724, USA

    Daniel W. Buster

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Contributions

D.Z. carried out and analysed most experiments using Drosophila S2 and human cells. D.Z. also purified and carried out the initial in vitro characterization of rDm-Kat60. K.D.G. carried out most experiments with Drosophila D17 under the direction of S.L.R.; J.D.C. also carried out analyses in D17 cells. S.F.S. designed the automated tracking algorithm under the direction of A.M. and used it to track microtubules in live-cell movies provided by D.Z.; J.D.D-V. and J.L.R. carried out numerous in vitro assays to quantify the microtubule-severing and end-depolymerase activities of rDM-Kat-60. A.B.A., E.L. and T.R. carried out and analysed the electron microscopy under the direction of H.J.S. U.R. carried out KLP59D for the electron microscopy assay. D.W.B. helped in the design of many experiments in S2 cells and made the model figure shown in Supplementary Information. D.J.S. wrote the manuscript (with the help of all authors, but particularly S.L.R.) and coordinated the efforts of the multiple laboratories involved in this project.

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Correspondence to David J. Sharp.

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Zhang, D., Grode, K., Stewman, S. et al. Drosophila katanin is a microtubule depolymerase that regulates cortical-microtubule plus-end interactions and cell migration. Nat Cell Biol 13, 361–369 (2011). https://doi.org/10.1038/ncb2206

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