Draft Genome Sequences of Three Clinical Isolates of Tannerella forsythia Isolated from Subgingival Plaque from Periodontitis Patients in the United States

Stafford, Graham P.; Chaudhuri, Roy R.; Haraszthy, Violet I.; Friedrich, Valentin; Schäffer, Christina; Ruscitto, Angela; Honma, Kiyonobu; Sharma, Ashu

doi:10.1128/genomea.01286-16

Cited by 9 publications

(11 citation statements)

References 16 publications

(16 reference statements)

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“…The amplification and sequencing of various NulO biosynthesis genes indicated that UB4 possesses the same Leg biosynthesis gene locus as strain FDC 92A2 (Supplementary Figure 4). This was confirmed by an alignment of the FDC 92A2 genome with the recently published genome of strain UB4, which revealed a nucleotide identity of 99.8% in this cluster (Stafford et al in preparation). The aminotransferase genes pseC and legC were selected as targets for gene knockouts in the ATCC 43037 type strain and UB4, respectively.…”

Section: Resultssupporting

confidence: 63%

“…A homology search of C. jejuni NulO biosynthesis amino acid sequences against the genomes of strains FDC 92A2 (Genbank CP003191) and UB4 (Stafford et al in preparation) revealed major differences with respect to the genome of strain ATCC 43037. The putative flippase gene (Tanf_01180/BFO_1075) is identical in FDC 92A2, UB4 and the type strain along almost its entire length, however, alignment breaks down over the last 30 base pairs (Supplementary Figure 1).…”

Section: Resultsmentioning

confidence: 99%

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Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

et al. 2016

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Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium’s cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead. In contrast to the NulO in ATCC 43037, which has been previously identified as a Pse derivative (5-N-acetimidoyl-7-N-glyceroyl-3,5,7,9-tetradeoxy-l-glycero-l-manno-NulO), glycan analysis of strain UB4 performed in this study indicated a 350-Da, possibly N-glycolyl Leg (3,5,7,9-tetradeoxy-d-glycero-d-galacto-NulO) derivative with unknown C5,7 N-acyl moieties. We have expressed, purified and characterized enzymes of both NulO pathways to confirm these genes’ functions. Using capillary electrophoresis (CE), CE–mass spectrometry and NMR spectroscopy, our studies revealed that Pse biosynthesis in ATCC 43037 essentially follows the UDP-sugar route described in Helicobacter pylori, while the pathway in strain FDC 92A2 corresponds to Leg biosynthesis in Campylobacter jejuni involving GDP-sugar intermediates. To demonstrate that the NulO biosynthesis enzymes are functional in vivo, we created knockout mutants resulting in glycans lacking the respective NulO. Compared to the wild-type strains, the mutants exhibited significantly reduced biofilm formation on mucin-coated surfaces, suggestive of their involvement in host-pathogen interactions or host survival. This study contributes to understanding possible biological roles of bacterial NulOs.

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Section: Resultssupporting

confidence: 63%

Section: Resultsmentioning

confidence: 99%

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

et al. 2016

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“…Mutants of T. forsythia ATCC 43037 (JUET00000000) and T. forsythia UB4 (FMMN01000000) with characterized defects in their cell surface protein glycosylation, affecting the terminal Pse (ATCC 43037) or Leg (UB4) residue, were available in our laboratory from a previous study . Briefly, T. forsythia ATCC 43037 ∆ pseC (coding for a dedicated aminotransferase from the Pse biosynthesis pathway) and T. forsythia UB4 ∆ legC (coding for a dedicated aminotransferase from the Leg biosynthesis pathway) mutants were constructed by chromosomal insertion of a gene knockout cassette consisting of an erythromycin resistance gene flanked by homologous upstream and downstream regions, ~1000 bp, each.…”

Section: Methodsmentioning

confidence: 99%

Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

Bloch¹,

Thurnheer

Murakami

et al. 2017

Molecular Oral Microbiology

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SummaryAs a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. The bacterium has a characteristic cell surface (S‐) layer modified with a unique O‐glycan. Both the S‐layer and the O‐glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. The influence of the T. forsythia S‐layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony‐forming unit counts and quantitative real‐time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S‐layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. This suggests a selective role for the glycosylated T. forsythia S‐layer in the positioning of this species within the biofilm, its co‐localization with P. gingivalis, and the prevalence of C. rectus. These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm.

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“…In this study, T. forsythia wild‐type strains ATCC 43037 (type strain, American Type Culture Collection, Manassas, VA) and T. forsythia UB4 as well as defined mutants thereof were cultured anaerobically in brain‐heart infusion (BHI) broth (Oxoid, Basingstoke, UK) and BHI agar plates, supplemented with N ‐acetylmuramic acid and horse serum as described previously . Mutants with characterized defects in protein glycosylation included T. forsythia ATCC 43037 ∆ pseC (deficient in Pse), T. forsythia ATCC 43037 ∆wecC (deficient in a trisaccharide glycan branch including Pse) and T. forsythia UB4 ∆ legC (deficient in Leg).…”

Section: Methodsmentioning

confidence: 99%

Immune response profiling of primary monocytes and oral keratinocytes to different Tannerella forsythia strains and their cell surface mutants

Bloch

Zwicker

Bostancı

et al. 2018

Molecular Oral Microbiology

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The oral pathogen Tannerella forsythia possesses a unique surface (S-) layer with a complex O-glycan containing a bacterial sialic acid mimic in the form of either pseudaminic acid or legionaminic acid at its terminal position. We hypothesize that different T. forsythia strains employ these stereoisomeric sugar acids for interacting with the immune system and resident host tissues in the periodontium. Here, we show how T. forsythia strains ATCC 43037 and UB4 displaying pseudaminic acid and legionaminic acid, respectively, and selected cell surface mutants of these strains modulate the immune response in monocytes and human oral keratinocytes (HOK) using a multiplex immunoassay. When challenged with T. forsythia, monocytes secrete proinflammatory cytokines, chemokines and vascular endothelial growth factor (VEGF) with the release of interleukin-1β (IL-1β) and IL-7 being differentially regulated by the two T. forsythia wild-type strains. Truncation of the bacteria's O-glycan leads to significant reduction of IL-1β and regulates macrophage inflammatory protein-1. HOK infected with T. forsythia produce IL-1Ra, chemokines and VEGF. Although the two wild-type strains elicit preferential immune responses for IL-8, both truncation of the O-glycan and deletion of the S-layer result in significantly increased release of IL-8, granulocyte-macrophage colony-stimulating factor and monocyte chemoattractant protein-1. Through immunofluorescence and confocal laser scanning microscopy of infected HOK we additionally show that T. forsythia is highly invasive and tends to localize to the perinuclear region. This indicates, that the T. forsythia S-layer and attached sugars, particularly pseudaminic acid in ATCC 43037, contribute to dampening the response of epithelial tissues to initial infection and hence play a pivotal role in orchestrating the bacterium's virulence.

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Draft Genome Sequences of Three Clinical Isolates of Tannerella forsythia Isolated from Subgingival Plaque from Periodontitis Patients in the United States

Cited by 9 publications

References 16 publications

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

Tannerella forsythiastrains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

Immune response profiling of primary monocytes and oral keratinocytes to different Tannerella forsythia strains and their cell surface mutants

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