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Structural basis of Fic-mediated adenylylation

Nature Structural & Molecular Biology volume 17, pages 10041010 (2010) | Download Citation

Abstract

The Fic family of adenylyltransferases, defined by a core HPFx(D/E)GN(G/K)R motif, consists of over 2,700 proteins found in organisms from bacteria to humans. The immunoglobulin-binding protein A (IbpA) from the bacterial pathogen Histophilus somni contains two Fic domains that adenylylate the switch1 tyrosine residue of Rho-family GTPases, allowing the bacteria to subvert host defenses. Here we present the structure of the second Fic domain of IbpA (IbpAFic2) in complex with its substrate, Cdc42. IbpAFic2-bound Cdc42 mimics the GDI-bound state of Rho GTPases, with both its switch1 and switch2 regions gripped by IbpAFic2. Mutations disrupting the IbpAFic2–Cdc42 interface impair adenylylation and cytotoxicity. Notably, the switch1 tyrosine of Cdc42 is adenylylated in the structure, providing the first structural view for this post-translational modification. We also show that the nucleotide-binding mechanism is conserved among Fic proteins and propose a catalytic mechanism for this recently discovered family of enzymes.

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Acknowledgements

We are grateful to the University of California, San Diego X-ray facility staff and the Advanced Light Source (beam line 8.2.1) staff of Lawrence Berkeley National Laboratory for beam access and help with data collection, Z. Xu and X. Guo for critically reading the manuscript and members of the Dixon laboratory for helpful discussions. This work was supported by US National Institutes of Health grant AI060662 to J.E.D.

Author information

Affiliations

  1. Department of Pharmacology, University of California, San Diego, La Jolla, California, USA.

    • Junyu Xiao
    • , Carolyn A Worby
    •  & Jack E Dixon
  2. Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California, USA.

    • Carolyn A Worby
    •  & Jack E Dixon
  3. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA.

    • Carolyn A Worby
    •  & Jack E Dixon
  4. Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California, USA.

    • Seema Mattoo
    •  & Jack E Dixon
  5. Berkeley Center for Structural Biology, Lawrence Berkeley Laboratory, Berkeley, California, USA.

    • Banumathi Sankaran

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Contributions

J.X. performed most of the research; S.M. purified VopS protein; B.S. collected diffraction data for IbpAFic2-alone crystals; J.X., C.A.W., S.M. and J.E.D. designed the research, analyzed data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jack E Dixon.

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DOI

https://doi.org/10.1038/nsmb.1867

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