The Spore Coat Protein CotE Facilitates Host Colonization by Clostridium difficile

Hong, Huynh A.; Ferreira, William T.; Hosseini, Siamand; Anwar, Saba; Hitri, Krisztina; Wilkinson, Anthony J.; Vahjen, Wilfried; Zentek, Jürgen; Soloviev, Mikhail; Cutting, Simon M.

doi:10.1093/infdis/jix488

Cited by 31 publications

(31 citation statements)

References 36 publications

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“…In Syrian hamsters, C. difficile spores have been shown to germinate within the 1st h in the small intestine (43). However, spores have been shown to be involved in the first steps of colonization, through adhesion to the host mucus (9). This suggests that pdaA1 mutant spores may not be fully cleared from the gastrointestinal tract and therefore establish infection.…”

Section: Discussionmentioning

confidence: 99%

“…It has also been hypothesized that spore persistence in the gastrointestinal tract is a potential factor in the recurrences and relapses of CDI (2,8). Moreover, a recent study has linked CotE, a spore coat protein, with host colonization, describing the spore as responsible for the initial colonization by targeting and binding the host mucus (9). Through their characteristics and contribution in pathophysiology, spores play a major role in CDI.…”

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confidence: 99%

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N-Deacetylases required for muramic-δ-lactam production are involved in Clostridium difficile sporulation, germination, and heat resistance

Coullon

Rifflet

Wheeler

et al. 2018

Journal of Biological Chemistry

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Edited by Chris WhitfieldSpores are produced by many organisms as a survival mechanism activated in response to several environmental stresses. Bacterial spores are multilayered structures, one of which is a peptidoglycan layer called the cortex, containing muramic-␦lactams that are synthesized by at least two bacterial enzymes, the muramoyl-L-alanine amidase CwlD and the N-deacetylase PdaA. This study focused on the spore cortex of Clostridium difficile, a Gram-positive, toxin-producing anaerobic bacterial pathogen that can colonize the human intestinal tract and is a leading cause of antibiotic-associated diarrhea. Using ultra-HPLC coupled with high-resolution MS, here we found that the spore cortex of the C. difficile 630⌬erm strain differs from that of Bacillus subtilis. Among these differences, the muramic-␦lactams represented only 24% in C. difficile, compared with 50% in B. subtilis. CD630_14300 and CD630_27190 were identified as genes encoding the C. difficile N-deacetylases PdaA1 and PdaA2, required for muramic-␦-lactam synthesis. In a pdaA1 mutant, only 0.4% of all muropeptides carried a muramic-␦lactam modification, and muramic-␦-lactams were absent in the cortex of a pdaA1-pdaA2 double mutant. Of note, the pdaA1 mutant exhibited decreased sporulation, altered germination, decreased heat resistance, and delayed virulence in a hamster infection model. These results suggest a much greater role for muramic-␦-lactams in C. difficile than in other bacteria, including B. subtilis. In summary, the spore cortex of C. difficile contains lower levels of muramic-␦-lactams than that of B. subtilis, and PdaA1 is the major N-deacetylase for muramic-␦-lactam biosynthesis in C. difficile, contributing to sporulation, heat resistance, and virulence.Clostridium difficile is a Gram-positive, spore-forming, toxin-producing anaerobic bacterium that can colonize the intes-

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

N-Deacetylases required for muramic-δ-lactam production are involved in Clostridium difficile sporulation, germination, and heat resistance

Coullon

Rifflet

Wheeler

et al. 2018

Journal of Biological Chemistry

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“…Along with this set of BclA-like collagens, CotE is also predicted from our assembly where it is possible that it might similarly be involved in a multi-component attachment process. The CotE protein is also thought to be involved in the colonisation of the gut by C. difficile through C-terminal binding and degradation of mucins [87]. Glycosylated mucins on the nematode cuticle are implicated as the target in the ‘Velcro’ model of attachment [12].…”

Section: Discussionmentioning

confidence: 99%

De novoassembly of thePasteuria penetransgenome reveals high plasticity, host dependency, and BclA-like collagens

Orr¹,

Mauchline²,

Cock³

et al. 2018

Preprint

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12Pasteuria penetrans is a gram-positive endospore forming bacterial parasite of Meloidogyne spp. the 13 most economically damaging genus of plant parasitic nematodes globally. The obligate antagonistic 14 nature of P. penetrans makes it an attractive candidate biological control agent. However, deployment 15 of P. penetrans for this purpose is inhibited by a lack of understanding of its metabolism and the 16 molecular mechanics underpinning parasitism of the host, in particular the initial attachment of the 17 endospore to the nematode cuticle. Several attempts to assemble the genomes of species within this 18 genus have been unsuccessful. Primarily this is due to the obligate parasitic nature of the bacterium 19which makes obtaining genomic DNA of sufficient quantity and quality which is free from 20 contamination challenging. Taking advantage of recent developments in whole genome amplification, 21 long read sequencing platforms, and assembly algorithms, we have developed a protocol to generate 22 large quantities of high molecular weight genomic DNA from a small number of purified endospores. 23We demonstrate this method via genomic assembly of P. penetrans. This assembly reveals a reduced 24 genome of 2.64Mbp estimated to represent 86% of the complete sequence; its reduced metabolism 25 reflects widespread reliance on the host and possibly associated organisms. Additionally, apparent 26 expansion of transposases and prediction of partial competence pathways suggest a high degree of 27 genomic plasticity. Phylogenetic analysis places our sequence within the Bacilli, and most closely 28 related to Thermoactinomyces species. Seventeen predicted BclA-like proteins are identified which 29 may be involved in the determination of attachment specificity. This resource may be used to develop 30 in vitro culture methods and to investigate the genetic and molecular basis of attachment specificity. 31 32 33

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“…To test this hypothesis, we assessed the localization patterns of mCherry fusions to the basement layer proteins, SpoIVA (IVA) and SipL, and the outer coat protein, CotE, in wildtype, ∆IIM, and ∆IIP strains. We chose to analyze SpoIVA and SipL because they are required for polymerized coat to localize around the forespore in C. difficile , while CotE is a surface-accessible coat protein (Permpoonpattana et al, 2013;Hong et al, 2017) that we have shown mislocalizes to coat 'beards' in a IIQ mutant (Fimlaid et al, 2015b). The ∆IIM and ∆IIP strain backgrounds were chosen because they exhibit varying degrees of engulfment defects: ∆IIM has subtle defects in engulfment and coat localization in TEM analyses, while ∆IIP exhibits the most severe engulfment and coat localization defects (Figs 4 and S6).…”

Section: Basement Layer Coat Proteins Adhere To the Forespore In Engumentioning

confidence: 99%

Differential requirements for conserved peptidoglycan remodeling enzymes during Clostridioides difficile spore formation

Ribis

Fimlaid

Shen

2018

Molecular Microbiology

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Spore formation is essential for the bacterial pathogen and obligate anaerobe, Clostridioides (Clostridium) difficile, to transmit disease. Completion of this process depends on the mother cell engulfing the developing forespore, but little is known about how engulfment occurs in C. difficile. In Bacillus subtilis, engulfment is mediated by a peptidoglycan degradation complex consisting of SpoIID, SpoIIP and SpoIIM, which are all individually required for spore formation. Using genetic analyses, we determined the functions of these engulfment-related proteins along with the putative endopeptidase, SpoIIQ, during C. difficile sporulation. While SpoIID, SpoIIP and SpoIIQ were critical for engulfment, loss of SpoIIM minimally impacted C. difficile spore formation. Interestingly, a small percentage of ∆spoIID and ∆spoIIQ cells generated heat-resistant spores through the actions of SpoIIQ and SpoIID, respectively. Loss of SpoIID and SpoIIQ also led to unique morphological phenotypes: asymmetric engulfment and forespore distortions, respectively. Catalytic mutant complementation analyses revealed that these phenotypes depend on the enzymatic activities of SpoIIP and SpoIID, respectively. Lastly, engulfment mutants mislocalized polymerized coat even though the basement layer coat proteins, SpoIVA and SipL, remained associated with the forespore. Collectively, these findings advance our understanding of several stages during infectious C. difficile spore assembly.

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The Spore Coat Protein CotE Facilitates Host Colonization by Clostridium difficile

Abstract: Spores of Clostridium difficile carry a chitinase enzyme whose function is to bind to mucin, leading to its degradation. We show in vivo that this is potentially an important element of colonization and virulence.

Cited by 31 publications

References 36 publications

N-Deacetylases required for muramic-δ-lactam production are involved in Clostridium difficile sporulation, germination, and heat resistance

N-Deacetylases required for muramic-δ-lactam production are involved in Clostridium difficile sporulation, germination, and heat resistance

De novoassembly of thePasteuria penetransgenome reveals high plasticity, host dependency, and BclA-like collagens

Differential requirements for conserved peptidoglycan remodeling enzymes during Clostridioides difficile spore formation

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