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Effective assembly of fimbriae in Escherichia coli depends on the translocation assembly module nanomachine

Nature Microbiology volume 1, Article number: 16064 (2016) Cite this article

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Abstract

Outer membrane proteins are essential for Gram-negative bacteria to rapidly adapt to changes in their environment. Intricate remodelling of the outer membrane proteome is critical for bacterial pathogens to survive environmental changes, such as entry into host tissues1–3. Fimbriae (also known as pili) are appendages that extend up to 2 μm beyond the cell surface to function in adhesion for bacterial pathogens, and are critical for virulence. The best-studied examples of fimbriae are the type 1 and P fimbriae of uropathogenic Escherichia coli, the major causative agent of urinary tract infections in humans. Fimbriae share a common mode of biogenesis, orchestrated by a molecular assembly platform called ‘the usher’ located in the outer membrane. Although the mechanism of pilus biogenesis is well characterized, how the usher itself is assembled at the outer membrane is unclear. Here, we report that a rapid response in usher assembly is crucially dependent on the translocation assembly module. We assayed the assembly reaction for a range of ushers and provide mechanistic insight into the β-barrel assembly pathway that enables the rapid deployment of bacterial fimbriae.

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Figure 1: Assembly of the usher FimD.
Figure 2: E. coli ΔtamA or ΔtamB mutants have defects in the assembly of FimD.
Figure 3: The TAM is required for the rapid deployment of fimbriae.
Figure 4: The TAM is required for the assembly of multiple different usher proteins.

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Acknowledgements

The authors thank R. Goode and O. Kleinfeld for proteomic analysis and R. Bamert, R. Dunstan, R. Grinter and E. Heinz for comments on the manuscript. The authors also thank S. Hultgren for providing strain UTI89-Ptet-fim. This work was supported by an NHMRC Program Grant (606788, to T.L. and R.A.S.) and an NHMRC Project grant (APP1042651, to M.A.S.). T.L. is an ARC Australian Laureate Fellow, MA.S. is an NHMRC Senior Research Fellow, H.H.S. is an ARC Super Science Fellow, M.J.B. is an NHMRC Biomedical Fellow, and I.D.H. is an ARC Laureate Postdoctoral Fellow.

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Authors and Affiliations

  1. Biomedicine Discovery Institute and Department of Microbiology, Infection & Immunity Program, Monash University, Clayton 3800, Australia

    Christopher Stubenrauch, Matthew J. Belousoff, Iain D. Hay, Hsin-Hui Shen & Trevor Lithgow

  2. Department of Materials Engineering, Monash University, Clayton 3800, Australia

    Hsin-Hui Shen

  3. Institute of Structural and Molecular Biology (ISMB), University College London and Birkbeck College, Malet Street, London WC1E 7HX, UK

    James Lillington & Gabriel Waksman

  4. School of Chemistry, Monash University, Clayton 3800, Australia

    Kellie L. Tuck

  5. School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia 4072, Australia

    Kate M. Peters, Minh-Duy Phan, Alvin W. Lo & Mark A. Schembri

  6. Department of Microbiology & Immunology, University of Melbourne, Parkville 3052, Australia

    Richard A. Strugnell

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Contributions

C.S., M.J.B., I.D.H., K.M.P., M.-D.P. and A.W.L. designed and carried out the analyses. C.S., M.J.B., I.D.H., J.L., M.A.S., H.H.S. and K.L.T. provided expertise for the analyses. M.A.S., M.J.B., I.D.H., J.L., R.A.S., G.W. and T.L. supervised experimental work and evaluated data. M.A.S., R.A.S., G.W. and T.L. evaluated the results and wrote the manuscript.

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Correspondence to Trevor Lithgow.

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Stubenrauch, C., Belousoff, M., Hay, I. et al. Effective assembly of fimbriae in Escherichia coli depends on the translocation assembly module nanomachine. Nat Microbiol 1, 16064 (2016). https://doi.org/10.1038/nmicrobiol.2016.64

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