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Structure-function relationship of CAP-Gly domains

Nature Structural & Molecular Biology volume 14, pages 959–967 (2007)Cite this article

Abstract

In all eukaryotes, CAP-Gly proteins control important cellular processes. The molecular mechanisms underlying the functions of CAP-Gly domains, however, are still poorly understood. Here we use the complex formed between the CAP-Gly domain of p150glued and the C-terminal zinc knuckle of CLIP170 as a model system to explore the structure-function relationship of CAP-Gly–mediated protein interactions. We demonstrate that the conserved GKNDG motif of CAP-Gly domains is responsible for targeting to the C-terminal EEY/F sequence motifs of CLIP170, EB proteins and microtubules. The CAP-Gly–EEY/F interaction is essential for the recruitment of the dynactin complex by CLIP170 and for activation of CLIP170. Our findings define the molecular basis of CAP-Gly domain function, including the tubulin detyrosination-tyrosination cycle. They further establish fundamental roles for the interaction between CAP-Gly proteins and C-terminal EEY/F sequence motifs in regulating complex and dynamic cellular processes.

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Figure 1: Binding studies and X-ray crystal structure of p150n–ClipZn2.
Figure 2: Binding interface of p150n–ClipZn2, and sequence conservation.
Figure 3: Sequence conservation and binding sites of CAP-Gly domains.
Figure 4: The C-terminal DDETF sequence segment of CLIP170 is required for dynactin recruitment to CLIP170-associated microtubules.
Figure 5: CAP-Gly domains as EEY/F motif–recognition domains.
Figure 6: Activation of CLIP170.
Figure 7: Schematic illustration of the protein-protein interaction network analyzed and discussed in the present study.

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Acknowledgements

We thank I. Hayashi for providing data before publication, F. Winkler and D. Kostrewa for support with the X-ray data processing, T. Güntert and Y. Bächtiger for technical assistance and C. Weirich for careful reading of the manuscript. X-ray data were collected at beamline X06SA of the Swiss Light Source. This work was supported by the Swiss National Science Foundation through grant 3100A0-109423 (to M.O.S.) and within the framework of the National Center of Competence in Research Structural Biology program.

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Author notes

  1. Anke Weisbrich and Srinivas Honnappa: These authors contributed equally to this work.

Authors and Affiliations

  1. Biomolecular Research, Structural Biology, Paul Scherrer Insititut, Villigen PSI, CH-5232, Switzerland

    Anke Weisbrich, Srinivas Honnappa, Rolf Jaussi, Daniel Frey & Michel O Steinmetz

  2. Biochemisches Institut der Universität Zürich, Winterthurerstrasse 190, Zürich, CH-8057, Switzerland

    Oksana Okhrimenko & Ilian Jelesarov

  3. Department of Cell Biology, Erasmus Medical Center, P.O. Box 2040, Rotterdam, 3000 CA, The Netherlands

    Anna Akhmanova

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  1. Anke Weisbrich
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Contributions

A.W. and S.H. designed and executed the cloning, protein purification and structure determination. R.J. designed and executed the cloning. O.O. and I.J. designed and executed the ITC experiments. D.F. purified the protein. A.A. designed and executed the fluorescence microscopy and FRET experiments and wrote the paper. M.O.S. designed the research, guided the project and wrote the paper.

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Correspondence to Michel O Steinmetz.

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Weisbrich, A., Honnappa, S., Jaussi, R. et al. Structure-function relationship of CAP-Gly domains. Nat Struct Mol Biol 14, 959–967 (2007). https://doi.org/10.1038/nsmb1291

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