Serotype-Converting Bacteriophage SfII Encodes an Acyltransferase Protein That Mediates 6-O-Acetylation of GlcNAc in Shigella flexneri O-Antigens, Conferring on the Host a Novel O-Antigen Epitope

Sun, Qiang; Knirel, Yuriy A.; Wang, Jianping; Luo, Xia; Senchenkova, Sof’ya N.; Lan, Ruiting; Shashkov, Alexander S.; Xu, Jianguo

doi:10.1128/jb.02009-14

Cited by 23 publications

(26 citation statements)

References 40 publications

(78 reference statements)

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“…Furthermore, phages with esterase tailspikes are likely to play an important role in maintaining the diversity in dense bacterial populations where their selective pressure leads to a dynamic coexistence of isogenic strains with different O-antigen acetylation variants. The importance of O-acetyl modifications of the O-antigen for host recognition and their impact on bacteriophage lifestyle were shown in experiments on serotype converting temperate phages infecting Shigella flexneri (Lindberg et al, 1978;Allison and Verma, 2000;Sun et al, 2014).…”

Section: G7c Alters Non-specific Phage Defense Of the Hostmentioning

confidence: 99%

Function of bacteriophage G7C esterase tailspike in host cell adsorption

Prokhorov

Riccio

Zdorovenko

et al. 2017

Molecular Microbiology

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Bacteriophages recognize and bind to their hosts with the help of receptor-binding proteins (RBPs) that emanate from the phage particle in the form of fibers or tailspikes. RBPs show a great variability in their shapes, sizes, and location on the particle. Some RBPs are known to depolymerize surface polysaccharides of the host while others show no enzymatic activity. Here we report that both RBPs of podovirus G7C - tailspikes gp63.1 and gp66 - are essential for infection of its natural host bacterium E. coli 4s that populates the equine intestinal tract. We characterize the structure and function of gp63.1 and show that unlike any previously described RPB, gp63.1 deacetylates surface polysaccharides of E. coli 4s leaving the backbone of the polysaccharide intact. We demonstrate that gp63.1 and gp66 form a stable complex, in which the N-terminal part of gp66 serves as an attachment site for gp63.1 and anchors the gp63.1-gp66 complex to the G7C tail. The esterase domain of gp63.1 as well as domains mediating the gp63.1-gp66 interaction is widespread among all three families of tailed bacteriophages.

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Section: G7c Alters Non-specific Phage Defense Of the Hostmentioning

confidence: 99%

Function of bacteriophage G7C esterase tailspike in host cell adsorption

Prokhorov

Riccio

Zdorovenko

et al. 2017

Molecular Microbiology

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“…Bacterial cells were washed and dried as described by Robbins & Uchida (1962). LPS was isolated with a yield of 6.5 % by the phenol/water procedure (Westphal & Jann, 1965), followed by dialysis of the extract without layer separation, and purification as described by Sun et al (2014).…”

Section: Methodsmentioning

confidence: 99%

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

Wang

Arbatsky

et al. 2016

Microbiology

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Franconibacter (Enterobacter, Cronobacter) pulveris bacteria share several typical characteristics with, and hence pose a challenge for the detection of, Cronobacter sakazakii, an emerging opportunistic pathogen, which can cause severe infections in neonates. A structurally variable O-specific polysaccharide (OPS) called O antigen provides the major basis for the typing of Gram-negative bacteria. We investigated the structure and genetics of the O antigen of F. pulveris G3872 (designated O1). An OPS was isolated by mild alkaline degradation of the LPS, whereas the same polysaccharide and its oligosaccharide fragments were obtained by mild acid degradation. Studies by sugar analysis and NMR spectroscopy showed that the OPS contained D-ribose, L-rhamnose (L-Rha) and a rarely occurring monosaccharide 4-deoxy-Darabino-hexose, and the OPS structure was established. The O-antigen gene cluster of F. pulveris G3872 between JUMPStart and gnd genes includes putative genes for glycosyltransferases, ATP-binding cassette (ABC)-transporter genes wzm and wzt, and genes for the synthesis of L-Rha, but no genes for the synthesis of 4-deoxy-D-arabino-hexose. A mutation test with the wzm gene confirmed that the OPS is synthesized and exported by the ABC-transporter-dependent pathway. A trifunctional transferase was suggested to catalyse formation of two glycosidic linkages and add a methyl group to the non-reducing end of the OPS to terminate the chain elongation. A carbohydrate-binding module that presumably recognizes the terminal methyl-modified monosaccharide was found at the C-terminus of Wzt. Primers specific for F. pulveris G3872 were designed based on the wzm gene, which has potential to be used for identification and detection of the O1 serogroup.

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“…All serotypes, except serotype 6, share a basic structure of the O antigen with a tetrasaccharide repeat composed of one N-acetylglucosamine (GlcNAc) and three rhamnose residues (Rha I to Rha III ), which define the group antigenic determinant 3,4 referred to as serotype Y (5). The addition and combination of glucosyl and/or O-acetyl and/or phosphoethanolamine (PEtN) residues to different sugars of this basic structure confer the immune response specificity as well as the diversity of serotypes defining type (I to VII) and group antigenic determinants (6,7,8, and E1037) (3), (see Fig. S1 in the supplemental material).…”

mentioning

confidence: 99%

“…S1 in the supplemental material). Recently, two novel O-acetylation groups, viz, an O-acetyl group added to Rha III at position 3 or 4 (3/4-O-acetylation) in S. flexneri serotypes 1a, 1b, 2a, 5a, Y, and 6 and one added to GlcNAc at position 6 (6-Oacetylation) in serotypes 2a, 3a, Y, and Yv, were identified, conferring to the host a novel antigenic determinant provisionally Љfactor 9 and 10Љ (corresponding to factors 9 and 10, respectively) (6)(7)(8).…”

mentioning

confidence: 99%

“…At present, seven of these bacteriophages have been identified (SfI, SfIC, SfII, Sf6, SfIV, SfV, and SfX) (9). Three genes arranged in a single operon are responsible for the addition of a glucosyl group at different positions of any of the four sugars in the backbone O antigen (type antigenic determinants I, II, IV, V, and VII and group antigenic determinant 7,8). Two of them, namely, gtrA and gtrB, are highly conserved and interchangeable, while gtr type is unique in each bacteriophage encoding gtrI, gtrIC, gtrII, gtrIV, gtrV, and gtrX genes, conferring serotype 1, 1c (proposed as 7), 2, 4, 5, and X, respectively.…”

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confidence: 99%

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PCR-Based Method for Shigella flexneri Serotyping: International Multicenter Validation

et al. 2019

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Shigella spp. are a leading cause of human diarrheal disease worldwide, with Shigella flexneri being the most frequently isolated species in developing countries. This serogroup is presently classified into 19 serotypes worldwide. We report here a multicenter validation of a multiplex-PCR-based strategy previously developed by Q. Sun, R. Lan, Y. Wang, A. Zhao, et al. (J Clin Microbiol 49:3766 -3770, 2011) for molecular serotyping of S. flexneri. This study was performed by seven international laboratories, with a panel of 71 strains (researchers were blind to their identity) as well as 279 strains collected from each laboratory's own local culture collections. This collaborative work found a high extent of agreement among laboratories, calculated through interrater reliability (IRR) measures for the PCR test that proved its robustness. Agreement with the traditional method (serology) was also observed in all laboratories for 14 serotypes studied, while specific genetic events could be responsible for the discrepancies among methodologies in the other 5 serotypes, as determined by PCR product sequencing in most of the cases. This work provided an empirical framework that allowed the use of this molecular method to serotype S. flexneri and showed several advantages over the traditional method of serological typing. These advantages included overcoming the problem of availability of suitable antisera in testing laboratories as well as facilitating the analysis of multiple samples at the same time. The method is also less time-consuming for completion and easier to implement in routine laboratories. We recommend that this PCR be adopted, as it is a reliable diagnostic and characterization methodology that can be used globally for laboratory-based shigella surveillance.

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Serotype-Converting Bacteriophage SfII Encodes an Acyltransferase Protein That Mediates 6-O-Acetylation of GlcNAc in Shigella flexneri O-Antigens, Conferring on the Host a Novel O-Antigen Epitope

Cited by 23 publications

References 40 publications

Function of bacteriophage G7C esterase tailspike in host cell adsorption

Function of bacteriophage G7C esterase tailspike in host cell adsorption

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

PCR-Based Method for Shigella flexneri Serotyping: International Multicenter Validation

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