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Nitrogen nutrition of E. coli in the mammalian intestine

ABSTRACT: Our focus of this study is to determine the differential expression of genes related to uptake and degradation of nitrogenous compounds in E. coli colonizing the mouse intestine as compared to E. coli grown in minimal medium in vitro. We colonized CD-1 male mice with E. coli MG1655 StrR wild type and extracted the total RNA from mouse cecal mucus (GSE217743). We also colonized CD-1 male mice with E. coli MG1655 ∆glnG StrR CamR and extracted the total RNA from mouse cecal mucus. We grew E. coli MG1655 StrR wild type in MOPS minimal medium containing high nitrogen (10 mM NH4Cl) and low nitrogen (3 mM NH4Cl) and extracted the total RNA from exponential phase and stationary phase culture. We also grew E. coli MG1655 ∆glnG StrR CamR in MOPS minimal medium containing low nitrogen (3 mM NH4Cl) and extracted the total RNA from exponential phase culture. The extracted RNA samples were sequenced, and differential expression analysis was done to determine the genes related to uptake and degradation of nitrogenous compounds that are important for E. coli colonization. Our results indication nitrogen is not limiting for E. coli in the intestine and genes related to uptake and degradation of several aminoacids, di- and tripeptides, aminosugars, ethanolamine, urea among others are upregualated in the mouse intestine as compared to invitro culture grown in high nitrogen conditions.

ORGANISM(S): Escherichia coli str. K-12 substr. MG1655

PROVIDER: GSE241658 | GEO | 2024/01/29

REPOSITORIES: GEO

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Project description:Transcription profiling of wild type E. coli MG1655, intestine-adapted E. coli MG1655star, and E. coli MG1655 flhD mutant grown on glucose, mannose, and mucus. We previously isolated a spontaneous mutant of E. coli K-12, strain MG1655, following passage through the streptomycin-treated mouse intestine, which has colonization traits superior to the wild-type parent strain (Leatham, et. al., 2005, Infect Immun 73:8039-49) The intestine-adapted strain (E. coli MG1655star) grew faster on several different carbon sources compared to the wild-type and was non-motile due to deletion of the flhD gene. To further characterize E. coli MG1655star, we used several high-throughput genomic approaches. Whole-genome pyrosequencing did not reveal any changes on its genome, aside from the deletion at the flhDC locus, that could explain the colonization advantage of E. coli MG1655star. Microarray analysis revealed modest, yet significant induction of catabolic gene systems across the genome in both E. coli MG1655star and the isogenic flhD mutant. Catabolome analysis with Biolog GN2 Microplates revealed an enhanced ability of both E. coli MG1655star and the isogenic flhD mutant to oxidize a wide variety of carbon sources. The results show that intestine-adapted E. coli MG1655star is more fit than the wild-type for intestinal colonization because loss of FlhD results in elevated expression of genes involved in carbon and energy metabolism, leading to more efficient carbon source utilization, which results in a higher population size in the intestine. Hence mutations that enhance metabolic efficiency confer a colonization advantage. Three strains were profiled: E. coli MG1655 wildtype, E. coli flhD, and an intestine adapted strain, MG1655star, derived from the wildtype and isolated from feces after 15 days in the streptomycin treated mouse intestine, which proved to be a better colonizer than the wildtype, were grown on MOPS minimal medium containing 0.2% glucose or mannose, or mucus (10 mg/ml) and RNA was extracted from logarithmic phase cultures, and also from mucus grown cells in late log phase.

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Nitrogen nutrition of E. coli in the mammalian intestine

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