Architecture and self-assembly of Clostridium sporogenes and Clostridium botulinum spore surfaces illustrate a general protective strategy across spore formers

Janganan, Thamarai K., Mullin, Nic, Dafis-Sagarmendi, Ainhoa, Brunt, Jason ORCID: https://orcid.org/0000-0002-7815-2165, Tzokov, Svetomir B., Stringer, Sandra, Moir, Anne, Chaudhuri, Roy R., Fagan, Robert P., Hobbs, Jamie K. and Bullough, Per A. (2020) Architecture and self-assembly of Clostridium sporogenes and Clostridium botulinum spore surfaces illustrate a general protective strategy across spore formers. mSphere, 5 (4). ISSN 2379-5042

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Abstract

Spores, the infectious agents of many Firmicutes, are remarkably resilient cell forms. Even distant relatives can have similar spore architectures although some display unique features; they all incorporate protective proteinaceous envelopes. We previously found that Bacillus spores can achieve these protective properties through extensive disulfide cross-linking of self-assembled arrays of cysteine-rich proteins. We predicted that this could be a mechanism employed by spore formers in general, even those from other genera. Here, we tested this by revealing in nanometer detail how the outer envelope (exosporium) in Clostridium sporogenes (surrogate for C. botulinum group I), and in other clostridial relatives, forms a hexagonally symmetric semipermeable array. A cysteine-rich protein, CsxA, when expressed in Escherichia coli, self-assembles into a highly thermally stable structure identical to that of the native exosporium. Like the exosporium, CsxA arrays require harsh “reducing” conditions for disassembly. We conclude that in vivo, CsxA self-organizes into a highly resilient, disulfide cross-linked array decorated with additional protein appendages enveloping the forespore. This pattern is remarkably similar to that in Bacillus spores, despite a lack of protein homology. In both cases, intracellular disulfide formation is favored by the high lattice symmetry. We have identified cysteine-rich proteins in many distantly related spore formers and propose that they may adopt a similar strategy for intracellular assembly of robust protective structures.<jats:p/>IMPORTANCE Bacteria such as those causing botulism and anthrax survive harsh conditions and spread disease as spores. Distantly related species have similar spore architectures with protective proteinaceous layers aiding adhesion and targeting. The structures that confer these common properties are largely unstudied, and the proteins involved can be very dissimilar in sequence. We identify CsxA as a cysteine-rich protein that self-assembles in a two-dimensional lattice enveloping the spores of several Clostridium species. We show that apparently unrelated cysteine-rich proteins from very different species can self-assemble to form remarkably similar and robust structures. We propose that diverse cysteine-rich proteins identified in the genomes of a broad range of spore formers may adopt a similar strategy for assembly.

Item Type: Article
Additional Information: Author Acknowledgements: We thank Chris Hill for help with EM of thin sections. All electron microscopy work was carried out in the University of Sheffield's Faculty of Science Electron Microscopy Facility. We also thank Paul Kemp-Russell and Simon Dixon, who fabricated the acoustic enclosure for AFM. J.K.H. and N.M. gratefully acknowledge the Imagine: Imaging Life initiative at the University of Sheffield and the EPSRC for financial support through its Program Grant scheme (Grant No. EP/I012060/1). P.A.B. and T.K.J. gratefully acknowledge the Wellcome Trust for financial support. A.D.-S. was in receipt of a White Rose BBSRC DTP studentship (grant no. BB/M011151/1).
Uncontrolled Keywords: anaerobes,atomic force microscopy,bacillus anthracis,bacillus cereus,bacillus subtilis,botulism,clostridium difficile,disulfide bonding,electron microscopy,nanomaterials,protein structure-function,sporulation,microbiology,molecular biology ,/dk/atira/pure/subjectarea/asjc/2400/2404
Faculty \ School: Faculty of Science > School of Biological Sciences
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Depositing User: LivePure Connector
Date Deposited: 19 Dec 2022 16:33
Last Modified: 03 Jan 2023 10:35
URI: https://ueaeprints.uea.ac.uk/id/eprint/90303
DOI: 10.1128/msphere.00424-20

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