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ABSTRACT: Comparison of mother-daughter vaginal Lactobacillus crispatus genome sequences

PROVIDER: PRJNA547620 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9215370_1.fastq.gz	Fastqsanger.gz
	SRR9215370_2.fastq.gz	Fastqsanger.gz
	SRR9215371_1.fastq.gz	Fastqsanger.gz
	SRR9215371_2.fastq.gz	Fastqsanger.gz
	SRR9215372_1.fastq.gz	Fastqsanger.gz

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Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

Project description:Bacterial vaginosis (BV), a common syndrome characterized by Lactobacillus-deficient vaginal microbiota, is associated with adverse health outcomes. BV often recurs after standard antibiotic therapy in part because antibiotics promote microbiota dominance by Lactobacillus iners instead of Lactobacillus crispatus, which has more beneficial health associations. Strategies to promote L. crispatus and inhibit L. iners are thus needed. We show that oleic acid (OA) and similar long-chain fatty acids simultaneously inhibit L. iners and enhance L. crispatus growth. These phenotypes require OA-inducible genes conserved in L. crispatus and related lactobacilli, including an oleate hydratase (ohyA) and putative fatty acid efflux pump (farE). FarE mediates OA resistance, while OhyA is robustly active in the vaginal microbiota and enhances bacterial fitness by biochemically sequestering OA in a derivative form only ohyA-harboring organisms can exploit. OA promotes L. crispatus dominance more effectively than antibiotics in an in vitro BV model, suggesting a metabolite-based treatment approach.

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Limosilactobacillus fermentum Limits Candida glabrata Growth by Ergosterol Depletion

Project description:Candida glabrata is a human-associated opportunistic fungal pathogen. It shares its niche with Lactobacillus spp. in the gastrointestinal and vaginal tract. In fact, Lactobacillus species are thought to competitively prevent Candida overgrowth. We investigated the molecular aspects of this antifungal effect by analyzing the interaction of C. glabrata strains with Limosilactobacillus fermentum. From a collection of clinical C. glabrata isolates, we identified strains with different sensitivities to L. fermentum in coculture. We analyzed the variation of their expression pattern to isolate the specific response to L. fermentum. C. glabrata-L. fermentum coculture induced genes associated with ergosterol biosynthesis, weak acid stress, and drug/chemical stress. L. fermentum coculture depleted C. glabrata ergosterol. The reduction of ergosterol was dependent on the Lactobacillus species, even in coculture with different Candida species. We found a similar ergosterol-depleting effect with other lactobacillus strains (Lactobacillus crispatus and Lactobacillus rhamosus) on Candida albicans, Candida tropicalis, and Candida krusei. The addition of ergosterol improved C. glabrata growth in the coculture. Blocking ergosterol synthesis with fluconazole increased the susceptibility against L. fermentum, which was again mitigated by the addition of ergosterol. In accordance, a C. glabrata Derg11 mutant, defective in ergosterol biosynthesis, was highly sensitive to L. fermentum. In conclusion, our analysis indicates an unexpected direct function of ergosterol for C. glabrata proliferation in coculture with L. fermentum.

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Asymmetrically inherited older proteins in budding yeast (synchronized culture method-2)

Project description:In budding yeast, damaged organelles and biomolecules are symmetrically segregated between mother and daughter cells, which can influence the replicating aging of mother cells. In some cases, older proteins are prone to be damaged and decline in their functions compared to newly synthesized proteins, likely to contribute the cellular aging. The purpose of this analysis is to uncover asymmetrically inherited older proteins, which are preferentially retained in mother cells during division into mother and rejuvenating daughter cells. To achieve this purpose, we conducted an originally developed strategy where mother and daughter cells are separated after just one cycle of synchronized culture, during which newly synthesized proteins are labeled with stable isotope amino acid. Synchronized culture was carried out using virgin cells, which have never produce daughter cells and enriched in G1 phase. We successfully identified more than 20 proteins whose older forms are asymmetrically inherited by mother cells, among which are proteins involved in the intracellular homeostasis of the proton concentration and the response to the stress of misfolded proteins.

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