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Genome sequencing of clinical Tannerella forsythia strains

ABSTRACT: Draft genome sequences of three clinical isolates of Tannerella forsythia isolated from sub-gingival plaque from periodontitis patients

PROVIDER: PRJEB15383 | ENA |

REPOSITORIES: ENA

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Tannerella forsythia intraoral infection and periodontitis in C57BL/6J mice

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Project description:The objectives of this investigation were to examine changes in the host transcriptional profiles during a Tannerella forsythia infection using a murine calvarial model of inflammation and bone resorption. T. forsythia ATCC 43037 was injected into the subcutaneous soft tissue over the calvaria of BALB/c mice for 3 days, after which the soft tissues and calvarial bones were excised. RNA was isolated from infected soft tissues and calvarial bones and analyzed for transcript profiles using Murine GeneChip® MG-MOE430A Affymetrix arrays to provide a molecular profile of the events that occur following infection of these tissues. We used mouse microarrays to detail the molecular profile of the events that occur following infection of calvarial and bone tissues and identified distinct classes of up- and down-regulated genes during this process.

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Project description:Tannerella forsythia is a Gram-negative oral pathogen known to possess an O-glycosylation system responsible for targeting multiple proteins associated with virulence at the three-residue motif (D)(S/T)(A/I/L/V/M/T). Multiple proteins have been identified to be decorated with a decasaccharide glycan composed of a poorly defined core plus a species-specific portion whose biosynthesis is largely characterized. To date, the glycosylation of mainly the two S-layer glycoproteins, TfsA and TfsB has been studied yet the true extent of glycosylation within this species has not been explored. In the present study we explore the glycoproteome of T. forsythia employing FAIMS based glycopeptide enrichment of a cell membrane fraction. We demonstrate that at least 13 glycans are utilized within the T. forsythia glycoproteome varying with respect to the presence of the three terminal sugars and the presence of fucose and digitoxose branches at the reducing end. To improve the localization of glycosylation events and enhance the detection of glycopeptides we applied trifluoromethanesulfonic acid treatment to allow the selective chemical cleavage of glycans. By reducing the chemical complexity of glycopeptides this dramatically improved the number of glycopeptides identified and our ability to localize glycosylation sites by ETD fragmentation leading to the identification of 312 putative glycosylation sites in 145 glycoproteins. Glycosylation site analysis revealed that glycosylation occurs on a much broader glycosylation motif than initially reported with glycosylation found at (D)(S/T)(A/I/L/V/M/T/S/C/G/F). The data confirm earlier predictions of hundreds of possible O-glycoproteins present in this organism.

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Project description:We investigated the association between subgingival bacterial profiles and gene expression patterns in gingival tissues of patients with periodontitis. A total of 120 patients undergoing periodontal surgery contributed with a minimum of two interproximal gingival papillae (range 2-4) from a maxillary posterior. Prior to tissue harvesting, subgingival plaque samples were collected from the mesial and distal aspects of each papilla. Gingival tissue RNA was extracted, reverse-transcribed, labeled, and hybridized with whole-genome microarrays (310 in total)

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