Structural and energetic basis of folded-protein transport by the FimD usher (original) (raw)

Nature volume 496, pages 243–246 (2013) Cite this article

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Abstract

Type 1 pili, produced by uropathogenic Escherichia coli, are multisubunit fibres crucial in recognition of and adhesion to host tissues1. During pilus biogenesis, subunits are recruited to an outer membrane assembly platform, the FimD usher, which catalyses their polymerization and mediates pilus secretion2. The recent determination of the crystal structure of an initiation complex provided insight into the initiation step of pilus biogenesis resulting in pore activation, but very little is known about the elongation steps that follow3. Here, to address this question, we determine the structure of an elongation complex in which the tip complex assembly composed of FimC, FimF, FimG and FimH passes through FimD. This structure demonstrates the conformational changes required to prevent backsliding of the nascent pilus through the FimD pore and also reveals unexpected properties of the usher pore. We show that the circular binding interface between the pore lumen and the folded substrate participates in transport by defining a low-energy pathway along which the nascent pilus polymer is guided during secretion.

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Protein Data Bank

Data deposits

The atomic coordinates and structure factors of FimD–FimC–FimF–FimG–FimH have been deposited in the Protein Data Bank under accession ID 4J3O.

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Acknowledgements

This work was funded by Medical Research Council grant 85602 to G.W. D.B. and E.P. are supported by grant P41 GM103533 from the National Institute of General Medical Studies at the US National Institutes of Health (NIH). S.J.H. was supported by grant AI029549 from the National Institute of Allergy and Infectious Disease at the NIH. We thank the staff of beamline ID23-1 at the European Synchrotron Radiation Facility, the staff of beamline IO2 at the Diamond Light source and A. Cole for technical assistance during data collection.

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Author notes

  1. Sebastian Geibel and Erik Procko: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Structural and Molecular Biology, University College London and Birkbeck College, Malet Street, London WC1E 7HX, UK,
    Sebastian Geibel & Gabriel Waksman
  2. The Howard Hughes Medical Institute and Department of Biochemistry, University of Washington, Seattle, 98195, Washington, USA
    Erik Procko & David Baker
  3. Center for Women’s Infectious Disease Research and Department of Molecular Microbiology, Washington University School of Medicine, St Louis, 63011, Missouri, USA
    Scott J. Hultgren

Authors

  1. Sebastian Geibel
  2. Erik Procko
  3. Scott J. Hultgren
  4. David Baker
  5. Gabriel Waksman

Contributions

S.G. and E.P. carried out the crystallographic and computational work, respectively. D.B. and G.W. supervised the work. S.G., E.P., D.B. and G.W. analysed the data. S.G., E.P., S.J.H., D.B. and G.W. wrote the paper.

Corresponding authors

Correspondence toDavid Baker or Gabriel Waksman.

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The authors declare no competing financial interests.

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Geibel, S., Procko, E., Hultgren, S. et al. Structural and energetic basis of folded-protein transport by the FimD usher.Nature 496, 243–246 (2013). https://doi.org/10.1038/nature12007

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Editorial Summary

Type 1 pili biogenesis

The FimD usher is a secretion and assembly nanomachine that catalyses the polymerization of the type 1 pili used by uropathogenic Escherichia coli to adhere to host cell surfaces, and mediates translocation of the pili across the outer membrane. In this study, Gabriel Waksman and colleagues present the crystal structure of the FimD usher, for the first time capturing the elongation step during pilus biogenesis.