Dataset Information

Proteome analysis of biofilm extracellular vesicles produced by Granulicatella adiacens and Granulicatella elegans.

ABSTRACT: Granulicatella adiacens and Granulicatella elegans are non-motile, non-spore-forming, facultatively anaerobic Gram-positive cocci. They are part of the normal oral flora but causes serious infections such as infective endocarditis. When oral bacteria accidentally enter the bloodstream due to transient tissue damage during dental procedures, they have the potential to attach to the endocardium or an equivalent surface of an indwelling prosthesis and cause infection. Many bacterial species produce extracellular vesicles (EVs) as a virulence strategy. The study of these EVs was initially limited to Gram-negative bacteria; however, their production was also observed in some Gram-positive bacteria. Recent research in our lab has found that G. adiacens produces abundant EVs in planktonic life style. However, EVs production by G. adiacens in biofilm mode is unknown. Therefore, the objective is to isolate and characterize EVs produced by this bacterium in biofilm mode. G. adiacens CCUG 27809 will be cultured on chocolate blood agar with 0.001% pyridoxal hydrochloride at 37 °C and 5% CO2 for 2 days. The EVs will be isolated using differential centrifugation and filtration protocol and then observed using electron scanning microscopy. Proteins in the biofilm EVs will be identified using mass spectrometry. Unraveling the EVs proteome composition of G. adiacens biofilms may help elucidate intricate virulence strategies of this species.

INSTRUMENT(S):

ORGANISM(S): Granulicatella Adiacens Granulicatella Elegans Bacteria

SUBMITTER: Maribasappa Karched

LAB HEAD: Maribasappa Karched, PhD

PROVIDER: PXD059541 | Pride | 2026-01-05

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	DR_MARIBASAPPA_PID211_GA_PROTEOMICS_ANALYSIS_02_12_2024.pdResult	Other
	DR_MARIBASAPPA_PID211_GE_PROTEOMICS_ANALYSIS_02_12_2024.pdResult	Other
	GA1ECV_BIOFILM_1.raw	Raw
	GA1ECV_BIOFILM_2.raw	Raw
	GA1ECV_BIOFILM_3.raw	Raw

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Publications

Differential Proteomic Analysis of Extracellular Vesicles Produced by Granulicatella adiacens in Biofilm vs. Planktonic Lifestyle.

Karched Maribasappa M Alkandari Sarah S

Dentistry journal 20251126 12

Background: Gram-positive bacteria, once considered incapable of producing extracellular vesicles (EVs) due to their thick peptidoglycan layer, are now known to secrete EVs that transport virulence factors and modulate host immunity. These EVs contribute to bacterial pathogenicity by facilitating biofilm formation, immune evasion, and inflammation. Granulicatella adiacens, an oral commensal associated with infective endocarditis, represents a clinically relevant model to study EV-m ...[more]

PMID: 41440315

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Project description:Francisella tularensis is a Gram-negative bacterium that is highly virulent in humans, causing the disease tularemia. F. novicida is closely related to F. tularensis and exhibits high virulence in mice, but is avirulent in healthy humans. A F. novicida-specific gene cluster (FTN0451-FTN0456) encodes two proteins with diguanylate cyclase (DGC) and phosphodiesterase (PDE) domains that modulate synthesis and degradation of cyclic di-GMP (cdGMP). No DGC or PDE containing proteins are present in the F. tularensis genome. F. novicida strains lacking either the two DGC/PDE genes cdgA and cdgB or the entire gene cluster (KKF457) are defective for biofilm formation. In addition, expression of CdgB or a heterologous DGC in KKF457 stimulated F. novicida biofilms, even in a strain lacking the biofilm regulator QseB. Genetic evidence suggests that CdgA is predominantly a PDE, while CdgB is predominantly a DGC. The F. novicida qseB strain has reduced cdgA and cdgB transcript levels, demonstrating a F. novicida biofilm signaling cascade that controls cdGMP levels. Interestingly, KKF457 with elevated cdGMP levels exhibited a decrease in intramacrophage replication and virulence in mice, as well as increased growth yield and biofilm formation in vitro. Microarray analyses revealed that cdGMP stimulated the transcription of a chitinase (ChiB) known to contribute to biofilm formation. Our results indicate that elevated cdGMP in F. novicida stimulates biofilm formation and inhibits virulence. We suggest that differences in human virulence between F. novicida and F. tularensis may be due in part to the absence of cdGMP signaling in F. tularensis. A whole genome NibleGen microarray was developed for multiple Francisella subspecies, using shared probes for common genes as well as unique probes for subspecies-specific genes. Included were probes representing the F. novicida U112 genome, with a maximum of 6 probes/gene. RNA was isolated from bacteria and purified as described above. The double-strand cDNA was generated and labeled with Cy3 according to the NimbleGen Gene Expression Array protocol (www.nimblegen.com), using SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and the DNA Labeling Kit (NimbleGen). Microarray results were analyzed using the ArrayStar software (DNASTAR).

Dataset Information

Proteome analysis of biofilm extracellular vesicles produced by Granulicatella adiacens and Granulicatella elegans.

Dataset's files

Publications

Differential Proteomic Analysis of Extracellular Vesicles Produced by <i>Granulicatella adiacens</i> in Biofilm vs. Planktonic Lifestyle.

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Dataset Information

Proteome analysis of biofilm extracellular vesicles produced by Granulicatella adiacens and Granulicatella elegans.

Dataset's files

Publications

Differential Proteomic Analysis of Extracellular Vesicles Produced by &lt;i&gt;Granulicatella adiacens&lt;/i&gt; in Biofilm vs. Planktonic Lifestyle.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Differential Proteomic Analysis of Extracellular Vesicles Produced by <i>Granulicatella adiacens</i> in Biofilm vs. Planktonic Lifestyle.