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PH domain of ELMO functions in trans to regulate Rac activation via Dock180

Nature Structural & Molecular Biology volume 11, pages 756762 (2004) | Download Citation

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Abstract

The members of the Dock180 superfamily of proteins are novel guanine nucleotide exchange factors (GEF) for Rho family GTPases and are linked to multiple biological processes from worms to mammals. ELMO is a critical regulator of Dock180, and the Dock180–ELMO complex functions as a bipartite GEF for Rac. We identified a mechanism wherein the PH domain of ELMO, by binding the Dock180–Rac complex in trans, stabilizes Rac in the nucleotide-free transition state. Mutagenesis studies reveal that this ELMO PH domain–dependent regulation is essential for the Dock180–ELMO complex to function in phagocytosis and cell migration. Genetic rescue studies in Caenorhabditis elegans using ELMO and its homolog CED-12 support the above observations in vivo. These data reveal a new mode of action of PH domains and a novel, evolutionarily conserved mechanism by which a bipartite GEF can activate Rac.

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Acknowledgements

We thank A. Bouton and J. Casanova for comments on the manuscript and V. Yajnik, as well as members of the Ravichandran laboratory, for helpful discussions. We also thank M. Matsuda for original Dock180 plasmids and I. Macara for the His-tagged bacterially produced Rac. This work was supported by a US National Institutes of Health (NIH) grant GM-64709 (to K.S.R.) and grants from the Swiss National Science Foundation, The Ernst Hadorn Foundation, and the European Union (FP5 project APOCLEAR) to M.O.H. C.G. was supported by an Infectious Diseases Training grant from the NIH.

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Affiliations

  1. Beirne Carter Center for Immunology Research, Department of Microbiology, University of Virginia, Charlottesville, Virginia 22908, USA.

    • Mingjian Lu
    • , Cynthia Grimsley
    • , Colin deBakker
    • , Enrico Brugnera
    • , Annie-Carole Tosello-Trampont
    • , Lisa B Haney
    •  & Kodi S Ravichandran
  2. Department of Pharmacology, University of Virginia, Charlottesville, Virginia 22908, USA.

    • Cynthia Grimsley
  3. Institute of Molecular Biology, University of Zurich, 8057 Zurich, Switzerland.

    • Jason M Kinchen
    • , Doris Klingele
    •  & Michael O Hengartner
  4. Department of Molecular Genetics and Microbiology, State University of New York at Stony Brook, Stony Brook, New York 11794, USA.

    • Jason M Kinchen
  5. Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.

    • Kent L Rossman
    •  & John Sondek

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Kodi S Ravichandran.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Dock WA/GE and Dock DSH3/GE mutants do not bind ELMO.

  2. 2.

    Supplementary Fig. 2

    W665A mutant of full-length ELMO is less active than wild-type ELMO in GEF assay.

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DOI

https://doi.org/10.1038/nsmb800

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