In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620

Broeker, N.K.; Kiele, Franziska; Casjens, Sherwood; Gilcrease, Eddie B.; Thalhammer, Anja; Koetz, Joachim; Barbirz, Stefanie

doi:10.3390/v10060289

Cited by 19 publications

(20 citation statements)

References 60 publications

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“…For P22 it was shown that particles open and eject about 80% of their DNA content when triggered with LPS (Andres et al, 2010). The kinetic profile of this process is conserved in other podovirus systems analyzed with the same method, but different LPS receptors (Broeker et al, 2018). This reflects that the rate-limiting particle opening step most probably involves rearrangements in conserved parts of the podovirus tail assembly.…”

Section: Bacteriophage P22 Omv Interactions Clearly Differ From Intermentioning

confidence: 84%

In vitro Analysis of O-Antigen-Specific Bacteriophage P22 Inactivation by Salmonella Outer Membrane Vesicles

et al. 2020

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Bacteriophages use a large number of different bacterial cell envelope structures as receptors for surface attachment. As a consequence, bacterial surfaces represent a major control point for the defense against phage attack. One strategy for phage population control is the production of outer membrane vesicles (OMVs). In Gram-negative host bacteria, O-antigen-specific bacteriophages address lipopolysaccharide (LPS) to initiate infection, thus relying on an essential outer membrane glycan building block as receptor that is constantly present also in OMVs. In this work, we have analyzed interactions of Salmonella (S.) bacteriophage P22 with OMVs. For this, we isolated OMVs that were formed in large amounts during mechanical cell lysis of the P22 S. Typhimurium host. In vitro, these OMVs could efficiently reduce the number of infective phage particles. Fluorescence spectroscopy showed that upon interaction with OMVs, bacteriophage P22 released its DNA into the vesicle lumen. However, only about one third of the phage P22 particles actively ejected their genome. For the larger part, no genome release was observed, albeit the majority of phages in the system had lost infectivity towards their host. With OMVs, P22 ejected its DNA more rapidly and could release more DNA against elevated osmotic pressures compared to DNA release triggered with protein-free LPS aggregates. This emphasizes that OMV composition is a key feature for the regulation of infective bacteriophage particles in the system.

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Section: Bacteriophage P22 Omv Interactions Clearly Differ From Intermentioning

confidence: 84%

In vitro Analysis of O-Antigen-Specific Bacteriophage P22 Inactivation by Salmonella Outer Membrane Vesicles

et al. 2020

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“…In fact, tailspike binding and cleavage activities seem to be necessary not only for positioning the phage particle on the OM, but also to prime its opening ([ 37 ] and references therein). In good correlation with this, it was shown that highly purified LPS can induce phage DNA ejection in vitro when incubated with virions having this type of tailspikes [ 60 , 61 , 62 ].…”

Section: Crossing the Bacterial Cell Envelope To Get Inside: A Talmentioning

confidence: 84%

Enzymes and Mechanisms Employed by Tailed Bacteriophages to Breach the Bacterial Cell Barriers

Fernandes

São-José

2018

Viruses

138

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Monoderm bacteria possess a cell envelope made of a cytoplasmic membrane and a cell wall, whereas diderm bacteria have and extra lipid layer, the outer membrane, covering the cell wall. Both cell types can also produce extracellular protective coats composed of polymeric substances like, for example, polysaccharidic capsules. Many of these structures form a tight physical barrier impenetrable by phage virus particles. Tailed phages evolved strategies/functions to overcome the different layers of the bacterial cell envelope, first to deliver the genetic material to the host cell cytoplasm for virus multiplication, and then to release the virion offspring at the end of the reproductive cycle. There is however a major difference between these two crucial steps of the phage infection cycle: virus entry cannot compromise cell viability, whereas effective virion progeny release requires host cell lysis. Here we present an overview of the viral structures, key protein players and mechanisms underlying phage DNA entry to bacteria, and then escape of the newly-formed virus particles from infected hosts. Understanding the biological context and mode of action of the phage-derived enzymes that compromise the bacterial cell envelope may provide valuable information for their application as antimicrobials.

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“…While the two genera differ in their O-antigen structure, they do share sugar residues (a rhamnose and a galactose) that may act as a common receptor for phage S144. Some variation of the O-antigen receptor is tolerated by some phages, such as the podovirus HK620 that cleaves the O-antigen of E. coli belonging to serogroup O18A both in the absence or presence of a branching glucose [62]. However, further experiments are needed to determine if O-antigen is the only receptor needed for S144 to eject its DNA in S. Muenster and C. sakazakii, as seen for phages HK620 and P22 [62,63], or if a secondary receptor is required for infection, as reported for phage T4 [64].…”

Section: Discussionmentioning

confidence: 99%

Phage S144, a New Polyvalent Phage Infecting Salmonella spp. and Cronobacter sakazakii

Gambino

Sørensen

Ahern

et al. 2020

IJMS

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Phages are generally considered species- or even strain-specific, yet polyvalent phages are able to infect bacteria from different genera. Here, we characterize the novel polyvalent phage S144, a member of the Loughboroughvirus genus. By screening 211 Enterobacteriaceae strains, we found that phage S144 forms plaques on specific serovars of Salmonella enterica subsp. enterica and on Cronobacter sakazakii. Analysis of phage resistant mutants suggests that the O-antigen of lipopolysaccharide is the phage receptor in both bacterial genera. The S144 genome consists of 53,628 bp and encodes 80 open reading frames (ORFs), but no tRNA genes. In total, 32 ORFs coding for structural proteins were confirmed by ESI-MS/MS analysis, whereas 45 gene products were functionally annotated within DNA metabolism, packaging, nucleotide biosynthesis and phage morphogenesis. Transmission electron microscopy showed that phage S144 is a myovirus, with a prolate head and short tail fibers. The putative S144 tail fiber structure is, overall, similar to the tail fiber of phage Mu and the C-terminus shows amino acid similarity to tail fibers of otherwise unrelated phages infecting Cronobacter. Since all phages in the Loughboroughvirus genus encode tail fibers similar to S144, we suggest that phages in this genus infect Cronobacter sakazakii and are polyvalent.

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In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620

Cited by 19 publications

References 60 publications

In vitro Analysis of O-Antigen-Specific Bacteriophage P22 Inactivation by Salmonella Outer Membrane Vesicles

In vitro Analysis of O-Antigen-Specific Bacteriophage P22 Inactivation by Salmonella Outer Membrane Vesicles

Enzymes and Mechanisms Employed by Tailed Bacteriophages to Breach the Bacterial Cell Barriers

Phage S144, a New Polyvalent Phage Infecting Salmonella spp. and Cronobacter sakazakii

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