Dataset Information

CotL, a new morphogenetic coat protein of Clostridium difficile

ABSTRACT: The strict anaerobe Clostridium difficile is an important nosocomial enteropathogen that has become the most common cause of antibiotic-associated diarrhea. The oxygen-resistant C. difficile spores play a central role in the infectious cycle, contributing to transmission, infection and recurrence. The spore surface layers, coat and exosporium, enable resistance of the spores to extreme physical and chemical stresses. Despite the critical importance of the spore in C. difficile infection, little is known about the mechanisms that orchestrate the assembly of its external layers. In this study, we identified and characterized a new C. difficile spore protein, named CotL, which is required for the assembly of the spore surface layers. The cotL gene was expressed in the mother cell compartment under the dual control of σE and σK. The CotL protein was localized in the C. difficile spore coat and cotL mutant spores had a major morphologic defect at the level of the coat/exosporium layers, as observed by transmission electron microscopy. Accordingly, spores of the cotL mutant contained a reduced amount of several proteins of the coat and exosporium, including CotB, CotE and CdeC. Additionally, cotL inactivation resulted in a defect in localization of late spore coat proteins in sporulating cells. Finally, spores of the cotL mutant were more sensitive to lysozyme and were impaired in germination. These defects are most likely associated with the structurally altered coat. Collectively, these results strongly suggest that CotL is a morphogenetic protein essential for the assembly of the spore coat in C. difficile.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Peptoclostridium Difficile (strain 630) (clostridium Difficile)

TISSUE(S): Prokaryotic Cell, Spore

SUBMITTER: Thibaut Douché

LAB HEAD: Bruno Dupuy

PROVIDER: PXD010359 | Pride | 2019-10-16

REPOSITORIES: Pride

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Publications

CotL, a new morphogenetic spore coat protein of Clostridium difficile.

Alves Feliciano Carolina C Douché Thibaut T Giai Gianetto Quentin Q Matondo Mariette M Martin-Verstraete Isabelle I Dupuy Bruno B

Environmental microbiology 20190221 3

The strict anaerobe Clostridium difficile is the most common cause of antibiotic-associated diarrhoea. The oxygen-resistant C. difficile spores play a central role in the infectious cycle, contributing to transmission, infection and recurrence. The spore surface layers, the coat and exosporium, enable the spores to resist physical and chemical stress. However, little is known about the mechanisms of their assembly. In this study, we characterized a new spore protein, CotL, which is required for ...[more]

PMID: 30556639

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Project description:Bacterial spores play an important role in disease initiation, transmission and persistence. The outermost spore layer, the exosporium, is important as it is the first point of contact between the spore and the environment and may be involved in spore adherence, protection and germination. Clostridium sordellii is a highly lethal, spore forming pathogen that causes soft-tissue infections, enteritis and toxic-shock syndrome. Despite the importance of C. sordellii spores in disease, spore proteins from this bacterium have not been defined or interrogated functionally. In this study, we identified the C. sordellii outer spore proteome and two of the identified proteins, CSA and CSB, were characterised using a genetic and phenotypic approach. Both proteins were essential for the correct formation and positioning of the C. sordellii spore coat and exosporium. The absence of CSA reduced sporulation levels and increased spore sensitivity to heat, sodium hydroxide and hydrochloric acid. By comparison, CSB was required for normal levels of spore adherence to cervical, but not vaginal, cells, with csb mutant spores having increased adherence properties. The establishment of a mouse infection model of the gastrointestinal tract for C. sordellii allowed the role of CSA and CSB to be interrogated in an infected host. Following the oral administration of spores to mice, the wild-type strain efficiently colonized the gastrointestinal tract, with the peak of bacterial numbers occurring at one day post-infection. Colonization was reduced by two logs at four days post-infection. By comparison, mice infected with the csb mutant did not show a reduction in bacterial numbers. The absence of CSB therefore allows the csb mutant to persist within the gastrointestinal tract. We conclude that C. sordellii outer spore proteins are important for the structural and functional integrity of spores, and for colonization and persistence during infection. Finally, Clostridium difficile, Bacillus cereus and Bacillus anthracis encode proteins with homology to CSA and CSB but these bacteria produce spores that are structurally dissimilar to those of C. sordellii, and the function of the proteins in these hosts is different to that in C. sordellii. These findings suggest that, despite their homology, spore proteins can have variable functions in different bacterial species which highlights the necessity of studying each spore protein in the cognate species from which it originates.

Dataset Information

CotL, a new morphogenetic coat protein of Clostridium difficile

Publications

CotL, a new morphogenetic spore coat protein of Clostridium difficile.

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