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A sporulation signature protease is required for assembly of the spore surface layers, germination and host colonization in Clostridioides difficile

Marini, Eleonora; Olivença, Carmen; Ramalhete, Sara; Aguirre, Andrea Martinez; Ingle, Patrick; Melo, Manuel N.; Antunes, Wilson; Minton, Nigel P.; Hernandez, Guillem; Cordeiro, Tiago N.; Sorg, Joseph A.; Serrano, Mónica; Henriques, Adriano O.

A sporulation signature protease is required for assembly of the spore surface layers, germination and host colonization in Clostridioides difficile Thumbnail


Authors

Eleonora Marini

Carmen Olivença

Sara Ramalhete

Andrea Martinez Aguirre

Patrick Ingle

Manuel N. Melo

Wilson Antunes

Nigel P. Minton

Guillem Hernandez

Tiago N. Cordeiro

Joseph A. Sorg

Mónica Serrano

Adriano O. Henriques



Abstract

A genomic signature for endosporulation includes a gene coding for a protease, YabG, which in the model organism Bacillus subtilis is involved in assembly of the spore coat. We show that in the human pathogen Clostridioidesm difficile, YabG is critical for the assembly of the coat and exosporium layers of spores. YabG is produced during sporulation under the control of the mother cell-specific regulators σE and σK and associates with the spore surface layers. YabG shows an N-terminal SH3-like domain and a C-terminal domain that resembles single domain response regulators, such as CheY, yet is atypical in that the conserved phosphoryl-acceptor residue is absent. Instead, the CheY-like domain carries residues required for activity, including Cys207 and His161, the homologues of which form a catalytic diad in the B. subtilis protein, and also Asp162. The substitution of any of these residues by Ala, eliminates an auto-proteolytic activity as well as interdomain processing of CspBA, a reaction that releases the CspB protease, required for proper spore germination. An in-frame deletion of yabG or an allele coding for an inactive protein, yabGC207A, both cause misassemby of the coat and exosporium and the formation of spores that are more permeable to lysozyme and impaired in germination and host colonization. Furthermore, we show that YabG is required for the expression of at least two σK-dependent genes, cotA, coding for a coat protein, and cdeM, coding for a key determinant of exosporium assembly. Thus, YabG also impinges upon the genetic program of the mother cell possibly by eliminating a transcriptional repressor. Although this activity has not been described for the B. subtilis protein and most of the YabG substrates vary among sporeformers, the general role of the protease in the assembly of the spore surface is likely to be conserved across evolutionary distance.

Citation

Marini, E., Olivença, C., Ramalhete, S., Aguirre, A. M., Ingle, P., Melo, M. N., Antunes, W., Minton, N. P., Hernandez, G., Cordeiro, T. N., Sorg, J. A., Serrano, M., & Henriques, A. O. (2023). A sporulation signature protease is required for assembly of the spore surface layers, germination and host colonization in Clostridioides difficile. PLoS Pathogens, 19(11), Article e1011741. https://doi.org/10.1371/journal.ppat.1011741

Journal Article Type Article
Acceptance Date Oct 9, 2023
Online Publication Date Nov 13, 2023
Publication Date Nov 13, 2023
Deposit Date Jun 20, 2025
Publicly Available Date Jun 24, 2025
Journal PLoS Pathogens
Print ISSN 1553-7366
Electronic ISSN 1553-7374
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 19
Issue 11
Article Number e1011741
DOI https://doi.org/10.1371/journal.ppat.1011741
Public URL https://nottingham-repository.worktribe.com/output/27599265
Publisher URL https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1011741

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Publisher Licence URL
https://creativecommons.org/licenses/by/4.0/

Copyright Statement
Copyright: © 2023 Marini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.





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