A Multivalent Adsorption Apparatus Explains the Broad Host Range of Phage phi92: a Comprehensive Genomic and Structural Analysis

Schwarzer, David; Buettner, Falk F. R.; Browning, C. H.; Nazarov, Sergey; Rabsch, Wolfgang; Bethe, Andrea; Oberbeck, Astrid; Bowman, Valorie D.; Stummeyer, Katharina; Mühlenhoff, Martina; Leiman, P.G.; Gerardy‐Schahn, Rita

doi:10.1128/jvi.00801-12

Cited by 107 publications

(127 citation statements)

References 82 publications

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“…Overall dimensions of the baseplate are 182 Å in the narrow part, 240 Å in the broad part below the sheath–tube, and 195 Å in height including the protruding tip complex (Fig 2A and B). The central tip protrudes ~40 Å below base of the cone and clearly resembles a central spike present in all baseplates of phages with contractile tails (Browning et al , 2012; Leiman & Shneider, 2012; Schwarzer et al , 2012; Harada et al , 2013; Shneider et al , 2013; Habann et al , 2014; Taylor et al , 2016). …”

Section: Resultsmentioning

confidence: 95%

“…8,309 baseplates and distal ends were identified after 2D classification, and 2,660 baseplates and 3,710 distal ends were further processed by RELION1.4 (Scheres, 2012) to obtain 3D density (Appendix Table S1). Based on available structural data of other contractile tail‐like structures (Schwarzer et al , 2012; Heymann et al , 2013; Shikuma et al , 2014; Ge et al , 2015; Nováček et al , 2016; Taylor et al , 2016), we applied a sixfold symmetry (C6) during refinement of the baseplate and the distal end. The final resolution was estimated to be on average 8.7 Å (local resolution distribution from 7.2 to 14.2 Å) for the baseplate and 7.5 Å (from 6.7 to 13.7 Å) for the distal end (Fig EV1A).…”

Section: Resultsmentioning

confidence: 99%

“…Our reconstruction also suggests that TssK protein may be involved in recruitment of certain effectors to the spike. Additionally, it is possible that some organisms evolved proteins that would serve as extensions for the wedge and would thus further increase the size of the baseplate cavity similarly to the variations in baseplate sizes seen in phages (Fu et al , 2011; Schwarzer et al , 2012; Hu et al , 2015; Büttner et al , 2016; Nováček et al , 2016; Taylor et al , 2016). …”

Section: Discussionmentioning

confidence: 99%

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Cryo‐ EM reconstruction of Type VI secretion system baseplate and sheath distal end

et al. 2017

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The bacterial Type VI secretion system (T6SS) assembles from three major parts: a membrane complex that spans inner and outer membranes, a baseplate, and a sheath–tube polymer. The baseplate assembles around a tip complex with associated effectors and connects to the membrane complex by TssK. The baseplate assembly initiates sheath–tube polymerization, which in some organisms requires TssA. Here, we analyzed both ends of isolated non‐contractile Vibrio cholerae sheaths by cryo‐electron microscopy. Our analysis suggests that the baseplate, solved to an average 8.0 Å resolution, is composed of six subunits of TssE/F2/G and the baseplate periphery is decorated by six TssK trimers. The VgrG/PAAR tip complex in the center of the baseplate is surrounded by a cavity, which may accommodate up to ~450 kDa of effector proteins. The distal end of the sheath, resolved to an average 7.5 Å resolution, shows sixfold symmetry; however, its protein composition is unclear. Our structures provide an important step toward an atomic model of the complete T6SS assembly.

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Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Cryo‐ EM reconstruction of Type VI secretion system baseplate and sheath distal end

et al. 2017

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“…The E. coli lytic myovirus phi92 has at least five different tail spikes and tail fiber proteins identified by cryoelectron microscopy, allowing it to infect a wide range of E. coli and Salmonella strains (54). Two different RBPs, LtfA and LtfB of the T5-like siphoviruses DT57C and DT571/2, recognize different O antigens, i.e., the O22 or O87 type and the O81 type, respectively (55).…”

Section: Discussionmentioning

confidence: 99%

The Presence of Two Receptor-Binding Proteins Contributes to the Wide Host Range of Staphylococcal Twort-Like Phages

Takeuchi

Osada

Azam

et al. 2016

Appl Environ Microbiol

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Thanks to their wide host range and virulence, staphylococcal bacteriophages (phages) belonging to the genus Twortlikevirus (staphylococcal Twort-like phages) are regarded as ideal candidates for clinical application for Staphylococcus aureus infections due to the emergence of antibiotic-resistant bacteria of this species. To increase the usability of these phages, it is necessary to understand the mechanism underlying host recognition, especially the receptor-binding proteins (RBPs) that determine host range. In this study, we found that the staphylococcal Twort-like phage ⌽SA012 possesses at least two RBPs. Genomic analysis of five mutant phages of ⌽SA012 revealed point mutations in orf103, in a region unique to staphylococcal Twort-like phages. Phages harboring mutated ORF103 could not infect S. aureus strains in which wall teichoic acids (WTAs) are glycosylated with ␣-N-acetylglucosamine (␣-GlcNAc). A polyclonal antibody against ORF103 also inhibited infection by ⌽SA012 in the presence of ␣-GlcNAc, suggesting that ORF103 binds to ␣-GlcNAc. In contrast, a polyclonal antibody against ORF105, a short tail fiber component previously shown to be an RBP, inhibited phage infection irrespective of the presence of ␣-GlcNAc. Immunoelectron microscopy indicated that ORF103 is a tail fiber component localized at the bottom of the baseplate. From these results, we conclude that ORF103 binds ␣-GlcNAc in WTAs, whereas ORF105, the primary RBP, is likely to bind the WTA backbone. These findings provide insight into the infection mechanism of staphylococcal Twort-like phages. IMPORTANCE Staphylococcus phages belonging to the genus Staphylococcus aureus, a Gram-positive coccus, is a commensal and pathogenic bacterium that causes opportunistic infections in humans and animals. Currently, antibiotic resistance in this species poses a threat to public health. Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired infections around the world (1). The emergence of such antibiotic-resistant bacteria requires development of alternatives to antibiotic-based therapies. One promising alternative is phage therapy, in which bacteriophages (phages) are used to treat bacterial infections (2, 3). In preclinical trials performed in mice, S. aureus infections (including MRSA infections) were successfully treated by use of phages (4). Therefore, phage therapy has attracted great interest as an alternative to antibiotics.Previously, we isolated the virulent staphylococcal phage ⌽SA012, which exerts lytic effects on a wide range of S. aureus isolates from bovine mastitis cases (5). Genomic analysis of ⌽SA012 revealed that it belongs to the genus Twortlikevirus, which contains the Staphylococcus phages Twort, K, and G, whereas the genus SPO1likevirus contains the Bacillus phage SPO1 as well as Listeria phages P100 and A511. Lactobacillus phage LP65 and Enterococcus phage ⌽EF24C were classified as orphans within the subfamily (6). Representatives of the two genera share the following features: they (i) hav...

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“…Bacteriophages are highly diverse and can be classified on the basis of their genome, morphology, and host specificity. Most phages are host specific, but some broad-host-range ones are able to invade different bacterial species and even different genera (19,20). Infection of bacterial cells occurs when the phage attaches itself to the host cell and injects viral and sometimes foreign DNA (RNA) into it in a penetration step.…”

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confidence: 99%

Bacteriophages Isolated from Chicken Meat and the Horizontal Transfer of Antimicrobial Resistance Genes

Shousha

Awaiwanont

Sofka

et al. 2015

Appl Environ Microbiol

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c Antimicrobial resistance in microbes poses a global and increasing threat to public health. The horizontal transfer of antimicrobial resistance genes was thought to be due largely to conjugative plasmids or transposons, with only a minor part being played by transduction through bacteriophages. However, whole-genome sequencing has recently shown that the latter mechanism could be highly important in the exchange of antimicrobial resistance genes between microorganisms and environments. The transfer of antimicrobial resistance genes by phages could underlie the origin of resistant bacteria found in food. We show that chicken meat carries a number of phages capable of transferring antimicrobial resistance. Of 243 phages randomly isolated from chicken meat, about a quarter (24.7%) were able to transduce resistance to one or more of the five antimicrobials tested into Escherichia coli ATCC 13706 (DSM 12242). Resistance to kanamycin was transduced the most often, followed by that to chloramphenicol, with four phages transducing tetracycline resistance and three transducing ampicillin resistance. Phages able to transduce antimicrobial resistance were isolated from 44% of the samples of chicken meat that we tested. The statistically significant (P ‫؍‬ 0.01) relationship between the presence of phages transducing kanamycin resistance and E. coli isolates resistant to this antibiotic suggests that transduction may be an important mechanism for transferring kanamycin resistance to E. coli. It appears that the transduction of resistance to certain antimicrobials, e.g., kanamycin, not only is widely distributed in E. coli isolates found on meat but also could represent a major mechanism for resistance transfer. The result is of high importance for animal and human health.

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A Multivalent Adsorption Apparatus Explains the Broad Host Range of Phage phi92: a Comprehensive Genomic and Structural Analysis

Cited by 107 publications

References 82 publications

Cryo‐ EM reconstruction of Type VI secretion system baseplate and sheath distal end

Cryo‐ EM reconstruction of Type VI secretion system baseplate and sheath distal end

The Presence of Two Receptor-Binding Proteins Contributes to the Wide Host Range of Staphylococcal Twort-Like Phages

Bacteriophages Isolated from Chicken Meat and the Horizontal Transfer of Antimicrobial Resistance Genes

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