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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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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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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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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DOI: https://doi.org/10.1038/nsmb1291
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