The Clostridium difficile Exosporium Cysteine (CdeC)-Rich Protein Is Required for Exosporium Morphogenesis and Coat Assembly

Barra-Carrasco, Jonathan; Olguín-Araneda, Valeria; Plaza-Garrido, Ángela; Miranda-Cárdenas, Camila; Cofré-Araneda, Glenda; Pizarro-Guajardo, Marjorie; Sarker, Mahfuzur R.; Paredes-Sabja, Daniel

doi:10.1128/jb.00369-13

Cited by 81 publications

(151 citation statements)

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“…In the case of rAdhE1, a molecular mass of 47 kDa was detected because only the N-terminal half of AdhE1 was cloned and expressed, while for rCdeC, only a small N-terminal domain (ϳ12 kDa) could be cloned. The appearance of multiple CdeC species of higher molecular masses is most probably due to extensive covalent cross-linking found in this cysteine-rich protein, as reported previously (25).…”

Section: Resultssupporting

confidence: 56%

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Mucosal Antibodies to the C Terminus of Toxin A Prevent Colonization of Clostridium difficile

et al. 2017

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Mucosal immunity is considered important for protection against Clostridium difficile infection (CDI). We show that in hamsters immunized with Bacillus subtilis spores expressing a carboxy-terminal segment (TcdA 26 -39 ) of C. difficile toxin A, no colonization occurs in protected animals when challenged with C. difficile strain 630. In contrast, animals immunized with toxoids showed no protection and remained fully colonized. Along with neutralizing toxins, antibodies to TcdA 26 -39 (but not to toxoids), whether raised to the recombinant protein or to TcdA 26 -39 expressed on the B. subtilis spore surface, cross-react with a number of seemingly unrelated proteins expressed on the vegetative cell surface or spore coat of C. difficile. These include two dehydrogenases, AdhE1 and LdhA, as well as the CdeC protein that is present on the spore. Anti-TcdA 26 -39 mucosal antibodies obtained following immunization with recombinant B. subtilis spores were able to reduce the adhesion of C. difficile to mucus-producing intestinal cells. This cross-reaction is intriguing yet important since it illustrates the importance of mucosal immunity for complete protection against CDI.

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Section: Resultssupporting

confidence: 56%

“…To identify proteins recognized by anti-TcdA 26 -39 PAbs, we used mass spectrometry (MS) analysis (see Table S1 in the supplemental material). In both vegetative cells and spore extracts, the ϳ100-kDa band was identified as AdhE1 (aldehyde alcohol dehydrogenase), and the 50-kDa sporespecific species was identified as CdeC (25).…”

Section: Resultsmentioning

confidence: 99%

Mucosal Antibodies to the C Terminus of Toxin A Prevent Colonization of Clostridium difficile

et al. 2017

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“…A cysteine-rich protein (CdeC) localized to the outermost spore layer was found to be important for the formation of the nap layer of the C. difficile spore and might be the anchoring protein linking the outermost layer to the spore coat (190). The CotA protein was shown to be important for proper assembly of the outermost spore layer, and the CotA, CotE (a bifunctional peroxiredoxin reductase and chitinase), CotF, and SodA (superoxide dismutase) proteins were found to be associated with the spore surface (185).…”

Section: Clostridium Difficilementioning

confidence: 99%

The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

Stewart

2015

Microbiol Mol Biol Rev

138

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SUMMARYMuch of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacteriumBacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present onB. subtilisspores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of other aspects of the spore. Because the bacterial spore surface is important for the spore's interactions with the environment, as well as being the site of interaction of the spore with the host's innate immune system in the case of spore-forming bacterial pathogens, the exosporium is worthy of continued investigation. Recent exosporium studies have focused largely on members of theBacillus cereusfamily, principallyBacillus anthracisandBacillus cereus. Our understanding of the composition of the exosporium, the pathway of its assembly, and its role in spore biology is now coming into sharper focus. This review expands on a 2007 review of spore surface layers which provided an excellent conceptual framework of exosporium structure and function (A. O. Henriques and C. P. Moran, Jr., Annu Rev Microbiol61:555–588, 2007,http://dx.doi.org/10.1146/annurev.micro.61.080706.093224). That review began a process of considering outer spore layers as an integrated, multilayered structure rather than simply regarding the outer spore components as independent parts.

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“…Spore formation is essential for C. difficile to survive exit from the host and transmit disease because its vegetative cells are exquisitely sensitive to oxygen (8). Furthermore, C. difficile spores are resistant to antibiotics (9), attacks from the host's immune system (10), and disinfectants commonly used in hospital settings due to their metabolic dormancy and intrinsic resistance properties (11)(12)(13)(14). Thus, understanding the mechanisms controlling C. difficile spore germination may have practical applications in the management of C. difficile infections as this knowledge may lead to new methods for preventing spore germination or efficiently promoting it to facilitate killing of the less resistant germinated spores.…”

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confidence: 99%

Characterization of the Dynamic Germination of Individual Clostridium difficile Spores Using Raman Spectroscopy and Differential Interference Contrast Microscopy

et al. 2015

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The Gram-positive spore-forming anaerobe Clostridium difficile is a leading cause of nosocomial diarrhea. Spores of C. difficile initiate infection when triggered to germinate by bile salts in the gastrointestinal tract. We analyzed germination kinetics of individual C. difficile spores using Raman spectroscopy and differential interference contrast (DIC) microscopy. Similar to Bacillus spores, individual C. difficile spores germinating with taurocholate plus glycine began slow leakage of a ϳ15% concentration of a chelate of Ca 2؉ and dipicolinic acid (CaDPA) at a heterogeneous time T 1 , rapidly released CaDPA at T lag , completed CaDPA release at T release , and finished peptidoglycan cortex hydrolysis at T lysis . T 1 and T lag values for individual spores were heterogeneous, but ⌬T release periods (T release ؊ T lag ) were relatively constant. In contrast to Bacillus spores, heat treatment did not stimulate spore germination in the two C. difficile strains tested. C. difficile spores did not germinate with taurocholate or glycine alone, and different bile salts differentially promoted spore germination, with taurocholate and taurodeoxycholate being best. Transient exposure of spores to taurocholate plus glycine was sufficient to commit individual spores to germinate. C. difficile spores did not germinate with CaDPA, in contrast to B. subtilis and C. perfringens spores. However, the detergent dodecylamine induced C. difficile spore germination, and rates were increased by spore coat removal although cortex hydrolysis did not follow T release , in contrast with B. subtilis. C. difficile spores lacking the cortex-lytic enzyme, SleC, germinated extremely poorly, and cortex hydrolysis was not observed in the few sleC spores that partially germinated. Overall, these findings indicate that C. difficile and B. subtilis spore germination exhibit key differences. IMPORTANCESpores of the Gram-positive anaerobe Clostridium difficile are responsible for initiating infection by this important nosocomial pathogen. When exposed to germinants such as bile salts, C. difficile spores return to life through germination in the gastrointestinal tract and cause disease, but their germination has been studied only with population-wide measurements. In this work we used Raman spectroscopy and DIC microscopy to monitor the kinetics of germination of individual C. difficile spores, the commitment of spores to germination, and the effect of germinant type and concentration, sublethal heat shock, and spore decoating on germination. Our data suggest that the order of germination events in C. difficile spores differs from that in Bacillus spores and provide new insights into C. difficile spore germination. C lostridium difficile is a Gram-positive, spore-forming, strictly anaerobic bacterium that has become a leading cause of antibiotic-associated diarrhea worldwide (1). Antibiotic treatment disrupts the normal colonic flora that typically suppresses C. difficile growth and allows ingested C. difficile spores to germinate, outgrow...

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The Clostridium difficile Exosporium Cysteine (CdeC)-Rich Protein Is Required for Exosporium Morphogenesis and Coat Assembly

Cited by 81 publications

References 42 publications

Mucosal Antibodies to the C Terminus of Toxin A Prevent Colonization of Clostridium difficile

Mucosal Antibodies to the C Terminus of Toxin A Prevent Colonization of Clostridium difficile

The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

Characterization of the Dynamic Germination of Individual Clostridium difficile Spores Using Raman Spectroscopy and Differential Interference Contrast Microscopy

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