Characteristics of the <i>Clostridium difficile</i> cell envelope and its importance in therapeutics

Kirk, Joseph A.; Banerji, Oishik; Fagan, Robert P.

doi:10.1111/1751-7915.12372

Cited by 55 publications

(62 citation statements)

References 116 publications

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“…The mechanisms by which bacteria build, maintain, and evolve their S-layers are important to human health and our ability to treat and respond to bacterial pathogens such as C. difficile, A. salmonicida, and B. anthracis 37–39 . Additionally, S-layers have been exploited as nanomaterials in a variety of applications 4,40 , including high-density organization and display of organic or inorganic molecules tethered to RsaA in particular 41,42 .…”

Section: Main Textmentioning

confidence: 99%

Continuous, Topologically Guided Protein Crystallization Controls Bacterial Surface Layer Self-Assembly

Comerci

Herrmann

Yoon

et al. 2019

Preprint

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19Bacteria assemble the cell envelope using localized enzymes to account for growth and 20 division of a topologically complicated surface 1-3 . However, a regulatory pathway has not been 21 identified for assembly and maintenance of the surface layer (S-layer), a 2D crystalline protein 22 coat surrounding the curved 3D surface of a variety of bacteria 4,5 . By specifically labeling, 23 performed time-resolved, super-resolution fluorescence imaging and single-molecule tracking 47 (SMT) of S-layer assembly on living C. crescentus cells. In C. crescentus, the S-layer is made of 48 a single 98 kDa SLP, RsaA, which accounts for around 30% of the cell's total protein synthesis 7 . 49RsaA, like other SLPs, self-assembles into crystalline sheets upon the addition of divalent 50 calcium (Ca 2+ ) in vitro [8][9][10][11][12] . Given the many fitness-related functions ascribed to crystalline 51 bacterial S-layers, we hypothesized that SLP self-assembly may play a role in generating the S-52 layer coat in vivo 4,6 . 53RsaA covers the cellular surface of C. crescentus by forming a 22 nm-repeat hexameric 54 crystal lattice and is non-covalently anchored to an ~18 nm thick LPS outer membrane 13-17 55 (Figure 1a,b). The surface topology of stalked and predivisional C. crescentus includes a 56 cylindrical stalk measuring roughly 100 nm in diameter while the crescentoid cell body 57 approaches 800 nm in width 18,19 (Figure 1d). This large variety of curved topologies guarantees 58 that crystal distortion and defects within the S-layer lattice structure are present, defects which 59 enable complete coverage of the bacterial surface 4,13,20,21 . We can use Gaussian curvature, the 60 product of the maximum and minimum curvatures at a given point, to quantify the cellular 61 topology 22 . Crystalline defects cluster at regions with high absolute values of Gaussian curvature 62 such as the cell poles and division plane, while grain boundaries occur where Gaussian curvature 63 approaches zero such as the cell body (Figure 1d) 20,21 . 64Specifically labeling the S-layer in vivo has proven difficult due to the SLP's life cycle 65 and functions, which include secretion, refolding, anchoring, and crystallization 11,16,17,23,24 . 66

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Section: Main Textmentioning

confidence: 99%

Continuous, Topologically Guided Protein Crystallization Controls Bacterial Surface Layer Self-Assembly

Comerci

Herrmann

Yoon

et al. 2019

Preprint

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“…As mentioned above, some species like Clostridium difficile, Clostridium botulinum, Clostridium perfringens and Clostridium tetani cause disease and infection in humans and animals (Dickson and Shevky, 1923;Orr and Reed, 1940;Hafiz and Oakley, 1976;Chang, 1978;Scaria et al, 2010;Smith et al, 2015;Lacey et al, 2016;Kirk et al, 2017). These species have been the focus of intensive study and strategies to combat them have been developed that involve the use of specific chemicals to interfere with cell wall functions, endospore germination and other essential functions (Best et al, 2010;Paul et al, 2010;Peck et al, 2011;Dembek et al, 2013;Brunt et al, 2014;Uzal et al, 2014;Lacey et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

The pangenome of the genus Clostridium

Udaondo

Duque

Ramos

2017

Environmental Microbiology

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The pangenome for the genus Clostridium sensu stricto, which was obtained using highly curated and annotated genomes from 16 species is presented; some of these cause disease, while others are used for the production of added-value chemicals. Multilocus sequencing analysis revealed that species of this genus group into at least two clades that include non-pathogenic and pathogenic strains, suggesting that pathogenicity is dispersed across the phylogenetic tree. The core genome of the genus includes 546 protein families, which mainly comprise those involved in protein translation and DNA repair. The GS-GOGAT may represent the central pathway for generating organic nitrogen from inorganic nitrogen sources. Glycerol and glucose metabolism genes are well represented in the core genome together with a set of energy conservation systems. A metabolic network comprising proteins/enzymes, RNAs and metabolites, whose topological structure is a non-random and scale-free network with hierarchically structured modules was built. These modules shed light on the interactions between RNAs, proteins and metabolites, revealing biological features of transcription and translation, cell wall biosynthesis, C1 metabolism and N metabolism. Network analysis identified four nodes that function as hubs and bottlenecks, namely, coenzyme A, HPr kinases, S-adenosylmethionine and the ribonuclease P-protein, suggesting pivotal roles for them in Clostridium.

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“…SLPs assemble to form planar sheets called S-layers on the surface of cells, where they are anchored usually through non-covalent interactions with other surface molecules such as lipopolysaccharide (LPS) in Gram-negative bacteria 2,6. S-layers act as the outermost permeability barriers protecting prokaryotic cells from extracellular attack and provide mechanical support to membranes 7.…”

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confidence: 99%

Structure of the hexagonal surface layer on Caulobacter crescentus cells

et al. 2017

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Many prokaryotic cells are encapsulated by a surface layer (S-layer) that consists of repeating units of S-layer proteins (SLPs). S-layers protect cells from the outside, provide mechanical stability, and also play roles in pathogenicity. In situ structural information on this highly abundant class of proteins is scarce, therefore atomic details of how S-layers are arranged on the surface of cells have remained elusive. Here, using purified C. crescentus’ sole SLP RsaA we obtained a 2.7 Å X-ray structure that shows the hexameric S-layer lattice. Next, we solved a 7.4 Å structure of the S-layer through electron cryotomography (cryo-ET) and sub-tomogram averaging of cell stalks. The X-ray structure was docked unambiguously into the cryo-ET map, resulting in a pseudo-atomic level description of the in vivo S-layer, which agrees completely with the atomic X-ray lattice model. This study spans different spatial scales from atoms to cells by combining X-ray crystallography with cryo-ET and sub-nanometre resolution sub-tomogram averaging. The cellular S-layer atomic structure shows that the S-layer is porous, with the largest gap dimension being 27 Å, and is stabilised by multiple Ca2+ ions bound near interfaces.

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Characteristics of the Clostridium difficile cell envelope and its importance in therapeutics

Cited by 55 publications

References 116 publications

Continuous, Topologically Guided Protein Crystallization Controls Bacterial Surface Layer Self-Assembly

Continuous, Topologically Guided Protein Crystallization Controls Bacterial Surface Layer Self-Assembly

The pangenome of the genus Clostridium

Structure of the hexagonal surface layer on Caulobacter crescentus cells

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