Project description:To investigate the effect of the Zn ionophore, PBT2, on the transcriptomic response of Fusobacterium nucleatum ATCC 25586, RNA was extracted from bacterial samples which had been treated as follows: Untreated (0.006% DMSO v/v), DMSO-Zn treated (0.006% DMSO v/v + 200 uM ZnSO4), or PBT2-Zn treated (0.125 ug/mL PBT2 + 200 uM ZnSO4) (DMSO was the vehicle control). RNA samples were collected at 0h, 0.5h, and 1h post challenge in biological triplicate and sequenced using Illumina HiSeq platform. We mapped sequences to the reference genome F. nucleatum subsp. nucleatum ATCC 25586 and performed DEseq2 analysis to determine differentially expressed genes across time and treatment.
Project description:Analysis of the region of interest for total protein samples or the outer membrane fraction of Fusobacterium nucleatum subsp. nucleatum ATCC 23726. The bacteria carried either an empty vector control, or a plasmid for the overexpression of the FoxI sRNA or its mutant version FoxI-3C.
Project description:Interspecies coaggregation promotes transcriptional changes of oral bacteria, contributing to the development of structurally balanced biofilms as well as oral diseases such as periodontitis. Streptococcus gordonii (S. gordonii) is an early colonizer of the oral cavity, and Fusobacterium nucleatum (F. nucleatum) may act as a bridge adhering to both early and late oral colonizers. These two species were commonly detected in healthy and periodontitis-diseased oral sites and could interact with immune cells such as macrophages. However, little research explored how intergeneric coaggregation affected transcriptional changes in S. gordonii and F. nucleatum subsp. polymorphum and how these gene changes might affect both species’ pathogenicity. The present study investigated transcriptional changes of both species in response to dual-species physical association using dual RNA-seq. Results indicated that after 30-min dual-species coaggregation, 148 genes were significantly up-regulated, and 124 genes were significantly down-regulated in S. gordonii. A total of 154 genes were significantly down-regulated, and 10 genes were significantly up-regulated in F. nucleatum subsp. polymorphum. A majority of up-regulated S. gordonii genes were involved in the biosynthesis and export of cell-wall proteins and the pathway of carbohydrate metabolism, and a group of down-regulated S. gordonii genes were associated with fatty acid biosynthesis and peptidoglycan biosynthesis. The transcriptome profiles indicated that the interspecies coaggregation led to a reduced level of DNA repair and lipopolysaccharides virulence in F. nucleatum subsp. polymorphum. The present study revealed that dual-species coaggregation induced a wide array of gene changes in S. gordonii and F. nucleatum subsp. polymorphum, enhancing S. gordonii’s adherence ability and attenuating F. nucleatum subsp. polymorphum's ability to produce LPS.
Project description:Fusobacterium nucleatum is a Gram-negative oral bacterial species associated with periodontal disease progression. As periodontal disease progresses, it is known F. nucleatum coaggregated with blood is frequently detected in the gingival crevice. However, it is largely unknown whether these interactions between F. nucleatum and blood induce a particular genetic response. We tested the cultures of F. nucleatum ATCC 25586 with or without blood in a semi-defined growth medium by microarray analysis and found 14 genes that were affected after only about 4hr. of adhered blood. Then, we selected 7 genes that changed significantly and tested these genes via real-time RT-PCR to confirm the validity of the microarray results. As a result, one amino sugar-binding protein on the membrane of F. nucleatum was especially expressed high via both microarray and real-time RT-PCR. Based upon these data, it appears that the protein on the F. nucleatum membrane binds and transfers the amino sugar only under blood conditions. This study aims to determine the effect of blood on the gene expression profiling of F. nucreatum. The study contains 2 separate experiments that both measure the cultures without or with blood. The samples with blood contain 9% and 33% blood.
Project description:Localization of Fusobacterium nucleatum in the placenta may be associated with pregnancy complications including preeclampsia (PE), but its specific pathobiology is unknown. Our aim was to analyze the effect of Fusobacterium nucleatum on HUVEC cells to further elucidate placental dysfunction in the context of Fusobacterium nucleatum infestation.