Formation of functional, non-amyloidogenic fibres by recombinant Bacillus subtilis TasA

Elliot Erskine, Ryan J. Morris, Marieke Schor, Chris Earl, Rachel M. C. Gillespie, Keith M. Bromley, Tetyana Sukhodub, Lauren Clark, Paul Fyfe, Louise C. Serpell, Nicola Stanley-Wall (Lead / Corresponding author), Cait E. MacPhee

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Abstract

Bacterial biofilms are communities of microbial cells encased within a self-produced polymeric matrix. In the Bacillus subtilis biofilm matrix the extracellular fibres of TasA are essential. Here a recombinant expression system allows interrogation of TasA, revealing that monomeric and fibre forms of TasA have identical secondary structure, suggesting that fibrous TasA is a linear assembly of globular units. Recombinant TasA fibres form spontaneously, and share the biological activity of TasA fibres extracted from B. subtilis, whereas a TasA variant restricted to a monomeric form is inactive and subjected to extracellular proteolysis. The biophysical properties of both native and recombinant TasA fibres indicate that they are not functional amyloid-like fibres. A gel formed by TasA fibres can recover after physical shear force, suggesting that the biofilm matrix is not static and that these properties may enable B. subtilis to remodel its local environment in response to external cues. Using recombinant fibres formed by TasA orthologues we uncover species variability in the ability of heterologous fibres to cross-complement the B. subtilis tasA deletion. These findings are indicative of specificity in the biophysical requirements of the TasA fibres across different species and/or reflect the precise molecular interactions needed for biofilm matrix assembly.
Original languageEnglish
Pages (from-to)897-913
Number of pages17
JournalMolecular Microbiology
Volume110
Issue number6
Early online date26 May 2018
DOIs
Publication statusPublished - Dec 2018

Keywords

  • TasA fibres
  • Bacillus subtilis
  • biofilm matrix
  • functional amyloid
  • amyloid-like fibres

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    Student Theses

    The structure and function of TasA in the Bacillus subtilis biofilm matrix

    Author: Erskine, E., 2019

    Supervisor: Stanley-Wall, N. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy

    Understanding the role of TapA in the formation of B. subtilis biofilms: a biochemical and genetic analysis

    Author: Earl, C., 2018

    Supervisor: Stanley-Wall, N. (Supervisor) & Coulthurst, S. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy

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    Stanley-Wall, Nicola

    Person: Academic

    Cite this

    Erskine, E., Morris, R. J., Schor, M., Earl, C., Gillespie, R. M. C., Bromley, K. M., Sukhodub, T., Clark, L., Fyfe, P., Serpell, L. C., Stanley-Wall, N., & MacPhee, C. E. (2018). Formation of functional, non-amyloidogenic fibres by recombinant Bacillus subtilis TasA. Molecular Microbiology, 110(6), 897-913. https://doi.org/10.1111/mmi.13985