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NSP-Cas protein structures reveal a promiscuous interaction module in cell signaling

Nature Structural & Molecular Biology volume 18, pages 1381–1387 (2011)Cite this article

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Abstract

Members of the novel SH2-containing protein (NSP) and Crk-associated substrate (Cas) protein families form multidomain signaling platforms that mediate cell migration and invasion through a collection of distinct signaling motifs. Members of each family interact via their respective C-terminal domains, but the mechanism of this association has remained enigmatic. Here we present the crystal structures of the C-terminal domain from the NSP protein BCAR3 and the complex of NSP3 with p130Cas. BCAR3 adopts the Cdc25-homology fold of Ras GTPase exchange factors, but it has a 'closed' conformation incapable of enzymatic activity. The structure of the NSP3–p130Cas complex reveals that this closed conformation is instrumental for interaction of NSP proteins with a focal adhesion-targeting domain present in Cas proteins. This enzyme-to-adaptor conversion enables high-affinity, yet promiscuous, interactions between NSP and Cas proteins and represents an unprecedented mechanistic paradigm linking cellular signaling networks.

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Figure 1: The BCAR3 C-terminal domain resembles a Cdc25-homology domain but adopts a closed conformation incapable of canonical GEF function.
Figure 2: The NSP3–p130Cas complex.
Figure 3: FAT domain of p130Cas is well defined and uses a previously unknown extended binding mode.
Figure 4: Analysis of NSP family–p130Cas interactions in vitro and in vivo.
Figure 5: NSP-Cas modules form class-specific yet promiscuous signaling nodes.

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Acknowledgements

We thank S. Snipas for protein sequencing, A. Bobkov for analytical ultracentrifugation and isothermal titration calorimetry and G. Salvesen for critical discussion of the manuscript. We also thank the Hope for a Cure Foundation for donation of equipment, J. Badger (DeltaG Technologies) for assistance in model evaluation, and the NKI Protein Facility for providing expression vectors. This work was supported by US National Institutes of Health (NIH) grants P01CA102583 and R01CA160457 to S.J.R. and E.B.P., R01CA116099 and P01HD025938 to E.B.P. and DOD-BCRP Fellowship BC100466 to P.D.M. Data collection at beamline X29 of the National Synchrotron Light Source was also supported by Biological and Environmental Research Department of Energy and the NIH National Center for Research Resources.

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Authors and Affiliations

  1. Program of Apoptosis and Cell Death Research, Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, USA

    Peter D Mace, Małgorzata K Dobaczewska, JeongEun J Lee & Stefan J Riedl

  2. Program of Signal Transduction, Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, California, USA

    Yann Wallez & Elena B Pasquale

  3. Department of Biology, Brookhaven National Laboratory, Upton, New York, USA

    Howard Robinson

  4. Pathology Department, University of California San Diego, San Diego, California, USA

    Elena B Pasquale

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  1. Peter D Mace
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Contributions

P.D.M. grew crystals, solved the crystal structures, designed and carried out in vitro experiments and wrote the manuscript. Y.W. designed, carried out and analyzed in vivo experiments, M.K.D. and J.J.L. expressed and purified proteins and grew initial NSP3–p130Cas crystals. H.R. carried out crystallographic data collection. S.J.R. and E.B.P. designed experiments, analyzed data and wrote the manuscript.

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Correspondence to Stefan J Riedl.

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Mace, P., Wallez, Y., Dobaczewska, M. et al. NSP-Cas protein structures reveal a promiscuous interaction module in cell signaling. Nat Struct Mol Biol 18, 1381–1387 (2011). https://doi.org/10.1038/nsmb.2152

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