Genetic Evidence for the Involvement of the S-Layer Protein Gene
 sap
 and the Sporulation Genes
 spo0A
 ,
 spo0B
 , and
 spo0F
 in Phage AP50c Infection of Bacillus anthracis

Plaut, Roger D.; Beaber, John W.; Zemansky, Jason; Kaur, Ajinder; George, Matroner; Biswas, Biswajit; Henry, Matthew; Bishop‐Lilly, Kimberly A.; Mokashi, Vishwesh; Hannah, Ryan M.; Pope, Robert K.; Read, Timothy D.; Stibitz, Scott; Calendar, Richard; Sozhamannan, Shanmuga

doi:10.1128/jb.00739-13

Cited by 29 publications

(38 citation statements)

References 65 publications

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“…It is clear that the S-layer is the cell surface receptor for all of the Avidocin-CDs described here and, by extension, the receptor for the bacteriophages from which the RBP-encoding genes were cloned. Although a Bacillus bacteriophage has been found to bind S-layer protein Sap (33), this is the first time a receptor for a C. difficile bacteriophage has been identified. Furthermore, our data imply that S-layer recognition is a common feature of bacteriophages that infect this species.…”

Section: Discussionmentioning

confidence: 95%

“…Despite a number of well-studied spore-forming organisms producing S-layers, including B. anthracis (33), S-layer biogenesis has not been reported previously to affect sporulation. There is currently no evidence to suggest that the SLPs are a structural part of the mature spore and the mechanisms by which the S-layer can influence sporulation are currently unknown.…”

Section: Discussionmentioning

confidence: 99%

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New class of precision antimicrobials redefines role of Clostridium difficile S-layer in virulence and viability

et al. 2017

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Avidocin-CDs are a new class of precision bactericidal agents that do not damage resident gut microbiota and are unlikely to promote the spread of antibiotic resistance. The precision killing properties result from the fusion of bacteriophage receptor binding proteins (RBPs) to a lethal contractile scaffold from an R-type bacteriocin. We recently described the prototypic Avidocin-CD, Av-CD291.2, that specifically kills C. difficile ribotype 027 strains and prevents colonization of mice. We have since selected two rare Av-CD291.2 resistant mutants of strain R20291 (RT027; S-layer cassette type-4, SCLT-4). These mutants have distinct point mutations in the slpA gene that result in an S-layer null phenotype. Reversion of the mutations to wild-type restored normal SLCT-4 S-layer formation and Av-CD291.2 sensitivity; however, complementation with other SCLT alleles did not restore Av-CD291.2 sensitivity despite restoring S-layer formation. Using newly identified phage RBPs, we constructed a panel of new Avidocin-CDs that kill C. difficile isolates in an SLCT-dependent manner, confirming the S-layer as the receptor in every case. In addition to bacteriophage adsorption, characterization of the S-layer null mutant also uncovered important roles for SlpA in sporulation, resistance to lysozyme and LL-37, and toxin production. Surprisingly, the S-layer-null mutant was found to persist in the hamster gut despite its completely attenuated virulence. Avidocin-CDs have significant therapeutic potential for the treatment and prevention of C. difficile Infection (CDI) given their exquisite specificity for the pathogen. Furthermore, the emergence of resistance forces mutants to trade virulence for continued viability and, therefore, greatly reduce their potential clinical impact.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

New class of precision antimicrobials redefines role of Clostridium difficile S-layer in virulence and viability

et al. 2017

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“…Thus, the SCWP of B. anthracis and pathogenic B. cereus strains functions as the structural determinant for bacteriophage-mediated lysis of bacteria by serving as ligand for the carbohydrate-binding domain of bacteriophage lysins (42). Furthermore, B. anthracis Sap functions as surface receptor for bacteriophage AP50c (110). …”

Section: Structure and Function Of The Secondary Cell Wall Polysacchamentioning

confidence: 99%

Assembly and Function of the Bacillus anthracis S-Layer

Missiakas

Schneewind

2017

Annu. Rev. Microbiol.

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Bacillus anthracis, the anthrax agent, is a member of the Bacillus cereus sensu lato group, which includes invasive pathogens of mammals or insects as well as nonpathogenic environmental strains. The genes for anthrax pathogenesis are located on two large virulence plasmids. Similar virulence plasmids have been acquired by other B. cereus strains and enable the pathogenesis of anthrax-like diseases. Among the virulence factors of B. anthracis is the S-layer-associated protein BslA, which endows bacilli with invasive attributes for mammalian hosts. BslA surface display and function are dependent on the bacterial S-layer, whose constituents assemble by binding to the secondary cell wall polysaccharide (SCWP) via S-layer homology (SLH) domains. B. anthracis and other pathogenic B. cereus isolates harbor genes for the secretion of S-layer proteins, for S-layer assembly, and for synthesis of the SCWP. We review here recent insights into the assembly and function of the S-layer and the SCWP.

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“…Of note, previous studies (Plaut et al . ) identified Sap as a possible receptor for AP50c, another B. anthracis ‐specific phage.…”

Section: Discussionmentioning

confidence: 99%

Detection ofBacillus anthracisspores from environmental water using bioluminescent reporter phage

Nguyen¹,

Makkar²,

Sharp³

et al. 2017

J Appl Microbiol

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Aims We investigated the ability of a temperate B. anthracis reporter phage (Wβ∷luxAB-2), which transduces bioluminescence to infected cells, to detect viable spores from deliberately contaminated environmental water samples. Methods and Results Environmental water was inoculated with spores and assayed with Wβ∷luxAB-2. Bioluminescent signals directly correlated with input phage and spore concentrations. A limit of detection of 101 and 102 CFU mL−1 within 8 hours was achieved from pond and lake water, respectively. Detection was greatly simplified by minimizing sample processing steps without spore extraction. The complex endogenous microbial flora and salt content of brackish water challenged the assay, extending the detection time to 12 hours for a sensitivity of 102 CFU mL−1. Phage-mediated bioluminescence was strictly dependent on bacterial physiology, being significantly reduced in mid/late log phase cells. This was shown to be due to an inability of the phage to adsorb. Conclusions The reporter phage Wβ∷luxAB-2 displays potential for simplified detection of viable spores from contaminated water samples within 12 hours.

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Genetic Evidence for the Involvement of the S-Layer Protein Gene sap and the Sporulation Genes spo0A , spo0B , and spo0F in Phage AP50c Infection of Bacillus anthracis

Cited by 29 publications

References 65 publications

New class of precision antimicrobials redefines role of Clostridium difficile S-layer in virulence and viability

New class of precision antimicrobials redefines role of Clostridium difficile S-layer in virulence and viability

Assembly and Function of the Bacillus anthracis S-Layer

Detection ofBacillus anthracisspores from environmental water using bioluminescent reporter phage

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