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Structural and mechanistic insights into the interaction between Rho and mammalian Dia

Nature volume 435, pages 513–518 (2005)Cite this article

Abstract

Formins are involved in a variety of cellular processes that require the remodelling of the cytoskeleton. They contain formin homology domains FH1 and FH2, which initiate actin assembly1,2. The Diaphanous-related formins form a subgroup that is characterized by an amino-terminal Rho GTPase-binding domain (GBD) and an FH3 domain, which bind somehow to the carboxy-terminal Diaphanous autoregulatory domain (DAD) to keep the protein in an inactive conformation3,4. Upon binding of activated Rho proteins, the DAD is released and the ability of the formin to nucleate and elongate unbranched actin filaments is induced. Here we present the crystal structure of RhoC in complex with the regulatory N terminus of mammalian Diaphanous 1 (mDia1) containing the GBD/FH3 region, an all-helical structure with armadillo repeats. Rho uses its ‘switch’ regions for interacting with two subdomains of GBD/FH3. We show that the FH3 domain of mDia1 forms a stable dimer and we also identify the DAD-binding site. Although binding of Rho and DAD on the N-terminal fragment of mDia1 are mutually exclusive, their binding sites are only partially overlapping. On the basis of our results, we propose a structural model for the regulation of mDia1 by Rho and DAD.

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Figure 1: Architecture and properties of mDiaN.
Figure 2: Overall structure of the mDiaN–Rho complex.
Figure 3: The Rho–mDiaN interaction.
Figure 4: Mutually exclusive binding of Rho and DAD to mDiaN.

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Acknowledgements

We thank P. Stege and D. Kuehlmann for expert technical assistance, T. Lorenz for the cloning of RhoC, I. Schlichting and W. Blankenfeldt for X-ray data collection, and the ESRF and DESY staff for support. A.W. thanks the Deutsche Forschungsgemeinschaft for financial support.

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Author notes

  1. R. Rose, M. Weyand and M. Lammers: *These authors contributed equally to this work

Authors and Affiliations

  1. Department of Structural Biology, Max-Planck-Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227, Dortmund, Germany

    R. Rose, M. Weyand, M. Lammers, T. Ishizaki, M. R. Ahmadian & A. Wittinghofer

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  1. R. Rose
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  2. M. Weyand
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Correspondence to A. Wittinghofer.

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Supplementary information

Supplementary Figure S1

Sequence alignment of Diaphanous-related formins. (PDF 12 kb)

Supplementary Figure S1 Legend

Legend for Supplementary Figure S1 (PDF 12 kb)

Supplementary Figure S2

Induction of stress-fibre formation in HeLa cells by RhoA mutants. (PDF 125 kb)

Supplementary Table S1

Binding constants for the tested combinations of mDiaN and Rho/DAD constructs as determined by stopped flow and ITC measurements. (PDF 26 kb)

Supplementary Table S2

Phasing Statistics. (PDF 24 kb)

Supplementary Table S3

Refinement Statistics. (PDF 45 kb)

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Rose, R., Weyand, M., Lammers, M. et al. Structural and mechanistic insights into the interaction between Rho and mammalian Dia. Nature 435, 513–518 (2005). https://doi.org/10.1038/nature03604

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