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Structural insights into host GTPase isoform selection by a family of bacterial GEF mimics

Nature Structural & Molecular Biology volume 16, pages 853–860 (2009)Cite this article

Abstract

The Escherichia coli type III effector Map belongs to a large family of bacterial virulence factors that activate host Rho GTPase signaling pathways through an unknown molecular mechanism. Here we report direct evidence that Map functions as a potent and selective guanine-nucleotide exchange factor (GEF) for Cdc42. The 2.3-Å structure of the Map–Cdc42 complex revealed that Map mimics the GEF strategy of the mammalian Dbl family but has a three-dimensional architecture that is nearly identical to the bacterial GEF Salmonella spp. SopE. A comparative analysis between human and bacterial GEFs revealed a previously uncharacterized pairing mechanism between Map and the variable β2-3 interswitch region of Cdc42. We propose a GTPase selection model that is experimentally validated by the preferential activation Rac1 and RhoA by the Shigella spp. effectors IpgB1 and IpgB2, respectively. These results significantly expand the repertoire of bacterial GEF mimics and unify a GEF selection mechanism for host GTPase substrates.

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Figure 1: Biochemical function and structure of Map in complex with Cdc42.
Figure 2: Compromise in in vitro and in vivo activity of Map by mutagenesis.
Figure 3: Structural comparison between Map, SopE and ITSN.
Figure 4: Structural comparison of the Map–Cdc42, SopE–Cdc42 and ISTN–Cdc42 complexes.
Figure 5: Structural determinants of Cdc42 isoform selection by Map.
Figure 6: A common GTPase selective pairing mechanism of Map family type III effectors.

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Acknowledgements

We would like to specifically thank J. Dixon (University of California, San Diego), M. Rosen and Kim Orth (University of Texas, Southwestern (UTSW)) for helpful discussion in preparation of this manuscript and for providing valuable reagents, and B. Kenny (Institute for Cell and Molecular Biosciences, Medical School, University of Newcastle) for wild-type EPEC2348/69 and EPECΔmap strains. S.E.S. and A.J.W. are supported by Texas ARP (grant 010019-0085-29007 to N.M.A.) and R.C.O is supported by the Welch foundation (grant I-1704 to N.M.A.). This work was supported by the Rita C. and William P. Clements Jr. endowment for scholars program (UTSW Medical Center) and by the Texas ARP grant 010019-0085-29007 to N.M.A., and by the Chinese Ministry of Science and Technology “863” grant no. 2008AA022305 and “973” grant no. 2006CB806704 to J.C.

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  1. Zhiwei Huang and Sarah E Sutton: These authors contributed equally to this work.

Authors and Affiliations

  1. National Institute of Biological Sciences, Zhong Guan Cun Life Science Park, Beijing, China

    Zhiwei Huang, Xiaojing Wu, Yingcai Feng & Jijie Chai

  2. Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Sarah E Sutton, Adam J Wallenfang, Robert C Orchard & Neal M Alto

  3. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, China

    Jijie Chai

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Contributions

N.M.A., J.C., Z.H. and S.E.S. designed the research; Z.H., S.E.S., N.M.A. and J.C. performed the research; A.J.W., R.C.O., Z.W. and Y.F. provided new reagents and analytical tools and performed experiments; N.M.A. and J.C. wrote the paper.

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Correspondence to Jijie Chai or Neal M Alto.

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Huang, Z., Sutton, S., Wallenfang, A. et al. Structural insights into host GTPase isoform selection by a family of bacterial GEF mimics. Nat Struct Mol Biol 16, 853–860 (2009). https://doi.org/10.1038/nsmb.1647

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