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The DCX-domain tandems of doublecortin and doublecortin-like kinase

Nature Structural & Molecular Biology volume 10, pages 324–333 (2003)Cite this article

Abstract

The doublecortin-like domains (DCX), which typically occur in tandem, are novel microtubule-binding modules. DCX tandems are found in doublecortin, a 360-residue protein expressed in migrating neurons; the doublecortin-like kinase (DCLK); the product of the RP1 gene that is responsible for a form of inherited blindness; and several other proteins. Mutations in the gene encoding doublecortin cause lissencephaly in males and the 'double-cortex syndrome' in females. We here report a solution structure of the N-terminal DCX domain of human doublecortin and a 1.5 Å resolution crystal structure of the equivalent domain from human DCLK. Both show a stable, ubiquitin-like tertiary fold with distinct structural similarities to GTPase-binding domains. We also show that the C-terminal DCX domains of both proteins are only partially folded. In functional assays, the N-terminal DCX domain of doublecortin binds only to assembled microtubules, whereas the C-terminal domain binds to both microtubules and unpolymerized tubulin.

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Figure 1: Solution structure of the N-terminal DCX domain of human doublecortin.
Figure 2: The crystal structure of the N-terminal DCX domain of the doublecortin-like kinase.
Figure 3: Comparison of molecular models of structurally related members of the ubiquitin superfamily.
Figure 4: Comparison of the stabilities of the isolated DCX domains and DCX tandems in doublecortin and DCLK.
Figure 5: Binding of doublecortin DCX domains to tubulin.
Figure 6: In vivo microtubule bundling by various doublecortin fragments.
Figure 7: Impact of two patient mutations on the structure and function of N-DCX.

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Acknowledgements

This research is supported by NIH grants to Z.S.D. and to C.A.W. A NATO Collaborative Link grant to Z.S.D. and J.O. is gratefully acknowledged. T.C. and D.K. are recipients of the Young Scholar Awards from the Foundation for Polish Science. J.O. is an International Scholar of the Howard Hughes Medical Institute. J.H.B. is supported by grants from the NIH and the Leukemia and Lymphoma Society. We thank N. Olekhnovich for excellent technical assistance, F. Abildgaard of NMRFAM for assistance with the collection of some of the NMR data, P. Sheffield for his contribution in the early phase of the project, A.V. Somlyo for helpful comments on the manuscript and L. Tamm for assistance with the tubulin polymerization experiments.

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  1. Myung Hee Kim, Tomasz Cierpicki, Urszula Derewenda and Zbigniew Dauter: The contributions of these authors were particularly important to this study.

Authors and Affiliations

  1. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, 22908-0736, Virginia, USA

    Myung Hee Kim, Tomasz Cierpicki, Urszula Derewenda, Yancho Devedjiev, Jacek Otlewski, John H. Bushweller & Zygmunt S. Derewenda

  2. Laboratory of Protein Engineering, Institute of Biochemistry & Molecular Biology, University of Wroclaw, Tamka 2, Wroclaw, 50-137, Poland

    Tomasz Cierpicki, Daniel Krowarsch & Jacek Otlewski

  3. Division of Neurogenetics, Department of Neurology, Beth Israel Deaconess Medical Center, Boston, 02115, Massachusetts, USA

    Yuanyi Feng & Christopher A. Walsh

  4. Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, NCI, Brookhaven National Laboratory, Upton, 11973, New York, USA

    Zbigniew Dauter

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Kim, M., Cierpicki, T., Derewenda, U. et al. The DCX-domain tandems of doublecortin and doublecortin-like kinase. Nat Struct Mol Biol 10, 324–333 (2003). https://doi.org/10.1038/nsb918

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