Prevalence of<i>Bacillus anthracis</i>-Like Organisms and Bacteriophages in the Intestinal Tract of the Earthworm<i>Eisenia fetida</i>

Schuch, Raymond; Pelzek, Adam J.; Kan, Sherry; Fischetti, Vincent A.

doi:10.1128/aem.02518-09

Cited by 35 publications

(39 citation statements)

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“…However, using ␥ as a diagnostic tool can lead to false positives because of the susceptibility of several Bacillus cereus strains to infection with this phage (10,11). Recent studies have shown that the host range of ␥ is less specific to B. anthracis than those of tectiviruses Wip1 and AP50 (1,12). For example, B. cereus ATCC 4342 is sensitive to infection with ␥ phage but not to infection with either Wip1 or AP50.…”

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confidence: 99%

“…These features indicate that Wip1 belongs to the family Tectiviridae, a relatively rare phage group with surprising structural similarity to and a proposed evolutionary lineage in common with the mammalian adenovirus (3,4). The Tectiviridae family consists of six isolates that infect Gram-negative bacteria, including PRD1 (5), and six that infect Gram-positive bacteria, including Bam35, Gil16, AP50, and Wip1 (1,(6)(7)(8). While PRD1 has been studied in detail, tectiviruses that infect Gram-positive bacteria are not as well characterized.…”

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confidence: 99%

“…Wip1 phage exhibits a very narrow host range and is highly specific to B. anthracis (1), the notorious biothreat agent and Grampositive bacterium that causes anthrax disease. The current gold standard for identifying suspected B. anthracis involves testing for ␥ phage sensitivity (9; http://www.bt.cdc.gov/agent/anthrax/faq/diagnosis .asp).…”

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confidence: 99%

“…Wip1 (for worm intestinal phage 1) is a recently identified phage that infects the pathogen Bacillus anthracis and was isolated from the intestinal tracts of Eisenia fetida worms (1). It is a tailless, double-stranded DNA phage possessing an internal lipid membrane beneath an icosahedral protein coat (2).…”

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Identification of a Ligand on the Wip1 Bacteriophage Highly Specific for a Receptor on Bacillus anthracis

et al. 2013

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bTectiviridae is a family of tailless bacteriophages with Gram-negative and Gram-positive hosts. The family model PRD1 and its close relatives all infect a broad range of enterobacteria by recognizing a plasmid-encoded conjugal transfer complex as a receptor. In contrast, tectiviruses with Gram-positive hosts are highly specific to only a few hosts within the same bacterial species. The cellular determinants that account for the observed specificity remain unknown. Here we present the genome sequence of Wip1, a tectivirus that infects the pathogen Bacillus anthracis. The Wip1 genome is related to other tectiviruses with Gram-positive hosts, notably, AP50, but displays some interesting differences in its genome organization. We identified Wip1 candidate genes for the viral spike complex, the structure located at the capsid vertices and involved in host receptor binding. Phage adsorption and inhibition tests were combined with immunofluorescence microscopy to show that the Wip1 gene product p23 is a receptor binding protein. His-p23 also formed a stable complex with p24, a Wip1 protein of unknown function, suggesting that the latter is involved with p23 in host cell recognition. The narrow host range of phage Wip1 and the identification of p23 as a receptor binding protein offer a new range of suitable tools for the rapid identification of B. anthracis.

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Identification of a Ligand on the Wip1 Bacteriophage Highly Specific for a Receptor on Bacillus anthracis

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“…The latter comprises bacteriophages that infect B. thuringiensis, such as Bam35, Gil01, and Gil16 (3); B. anthracis, such as AP50 and Wip1 (4,5); and B. cereus, such as Sole or Simila (6,7). Contrary to most temperate bacteriophages, members of this group are not integrative viruses; rather, they replicate as linear episomic plasmids during the lysogenic cycle (8), in which late phage functions are suppressed (9).…”

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Bam35 Tectivirus Intraviral Interaction Map Unveils New Function and Localization of Phage ORFan Proteins

Berjón-Otero

Lechuga

Mehla

et al. 2017

J Virol

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The family Tectiviridae comprises a group of tailless, icosahedral, membranecontaining bacteriophages that can be divided into two groups by their hosts, either Gram-negative or Gram-positive bacteria. While the first group is composed of PRD1 and nearly identical well-characterized lytic viruses, the second one includes more variable temperate phages, like GIL16 or Bam35, whose hosts are Bacillus cereus and related Gram-positive bacteria. In the genome of Bam35, nearly half of the 32 annotated open reading frames (ORFs) have no homologs in databases (ORFans), being putative proteins of unknown function, which hinders the understanding of their biology. With the aim of increasing knowledge about the viral proteome, we carried out a comprehensive yeast two-hybrid analysis of all the putative proteins encoded by the Bam35 genome. The resulting protein interactome comprised 76 unique interactions among 24 proteins, of which 12 have an unknown function. These results suggest that the P17 protein is the minor capsid protein of Bam35 and P24 is the penton protein, with the latter finding also being supported by iterative threading protein modeling. Moreover, the inner membrane transglycosylase protein P26 could have an additional structural role. We also detected interactions involving nonstructural proteins, such as the DNA-binding protein P1 and the genome terminal protein (P4), which was confirmed by coimmunoprecipitation of recombinant proteins. Altogether, our results provide a functional view of the Bam35 viral proteome, with a focus on the composition and organization of the viral particle.IMPORTANCE Tailless viruses of the family Tectiviridae can infect commensal and pathogenic Gram-positive and Gram-negative bacteria. Moreover, they have been proposed to be at the evolutionary origin of several groups of large eukaryotic DNA viruses and self-replicating plasmids. However, due to their ancient origin and complex diversity, many tectiviral proteins are ORFans of unknown function. Comprehensive protein-protein interaction (PPI) analysis of viral proteins can eventually disclose biological mechanisms and thus provide new insights into protein function unattainable by studying proteins one by one. Here we comprehensively describe intraviral PPIs among tectivirus Bam35 proteins determined using multivector yeast twohybrid screening, and these PPIs were further supported by the results of coimmunoprecipitation assays and protein structural models. This approach allowed us to propose new functions for known proteins and hypothesize about the biological role of the localization of some viral ORFan proteins within the viral particle that will be helpful for understanding the biology of tectiviruses infecting Gram-positive bacteria.KEYWORDS tectivirus, Bam35, yeast two-hybrid, intraviral interactome, ORFan, protein-protein interactions, PPIs

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Isolation, Culture, and Characterization of Bacteriophages

Pelzek

Schuch

Schmitz

et al. 2008

CP Essential Lab Tech

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Bacteriophages (phages) are viruses that infect bacteria. Phages are the most abundant biological entities on Earth, and have influential effects on every ecosystem. Phage genomes represent a vast gene pool from which bacteria can draw upon for rapid evolution. Since the co‐discovery of phages in 1915 by Frederick Twort and 1917 by Felix d'Herelle, their potential as subjects of laboratory research has been exploited to great effect. Phage research has resulted in important strides in biology, from the elucidation of the function of DNA and the discovery of messenger RNA to the study of basic molecular interactions and genetic regulation. These fascinatingly simple and diverse entities are also valuable as diagnostic tools, genetic screening vectors, and potential therapeutics. This article provides an overview of techniques essential for entering the world of bacteriophage study, and contains protocols for the propagation, maintenance, storage, and basic characterization of phages. Curr. Protoc. Essential Lab. Tech. 7:4.4.1‐4.4.33. © 2013 by John Wiley & Sons, Inc.

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Prevalence ofBacillus anthracis-Like Organisms and Bacteriophages in the Intestinal Tract of the EarthwormEisenia fetida

Abstract: Stable infection of

Cited by 35 publications

References 40 publications

Identification of a Ligand on the Wip1 Bacteriophage Highly Specific for a Receptor on Bacillus anthracis

Identification of a Ligand on the Wip1 Bacteriophage Highly Specific for a Receptor on Bacillus anthracis

Bam35 Tectivirus Intraviral Interaction Map Unveils New Function and Localization of Phage ORFan Proteins

Isolation, Culture, and Characterization of Bacteriophages

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