A chemically defined minimal medium for the optimal culture of Listeria

Phan‐Thanh, Luu; Gormon, Thierry

doi:10.1016/s0168-1605(96)01205-6

Cited by 85 publications

(89 citation statements)

References 10 publications

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“…Bacteria were grown at 37°C in BHI medium. When indicated, the chemically defined minimal medium IMM (25) (26). These cells were propagated in Eagle's minimal essential medium (MEM) containing 10% (v/v) fetal bovine serum (FBS), 4 mM L-glutamine, and nonessential amino acids in BioDish-XL plates (BD Biosciences, product 351040) to reach ϳ80% confluency (ϳ5.6 ϫ 10 7 epithelial cells).…”

Section: Methodsmentioning

confidence: 99%

“…This was of significance because it could allow us to perform biochemical studies requiring large amounts of peptidoglycan, technically difficult to obtain from intracellular bacteria. For this purpose, bacteria were grown at 37°C in the chemically defined medium IMM described to be optimal for growing Listeria (25). Examination at the electron microscope of bacteria grown in BHI and IMM media denoted drastic morphological differences in the cell wall.…”

Section: Characterization Of Cell Wall Proteome Of L Monocytogenes Gmentioning

confidence: 99%

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Association of ActA to Peptidoglycan Revealed by Cell Wall Proteomics of Intracellular Listeria monocytogenes

Portillo

Calvo

D'Orazio

et al. 2011

Journal of Biological Chemistry

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Listeria monocytogenes is a Gram-positive intracellular bacterial pathogen that colonizes the cytosol of eukaryotic cells. Recent transcriptomic studies have revealed that intracellular L. monocytogenes alter expression of genes encoding envelope components. However, no comparative global analysis of this cell wall remodeling process is yet known at the protein level. Here, we used high resolution mass spectrometry to define the cell wall proteome of L. monocytogenes growing inside epithelial cells. When compared with extracellular bacteria growing in a nutrient-rich medium, a major difference found in the proteome was the presence of the actin assembly-inducing protein ActA in peptidoglycan purified from intracellular bacteria. ActA was also identified in the peptidoglycan of extracellular bacteria growing in a chemically defined minimal medium. In this condition, ActA maintains its membrane anchoring domain and promotes efficient bacterial entry into nonphagocytic host cells. Unexpectedly, Internalin-A, which mediates entry of extracellular L. monocytogenes into eukaryotic cells, was identified at late infection times (6 h) as an abundant protein in the cell wall of intracellular bacteria. Other surface proteins covalently bound to the peptidoglycan, as Lmo0514 and Lmo2085, were detected exclusively in intracellular and extracellular bacteria, respectively. Altogether, these data provide the first insights into the changes occurring at the protein level in the L. monocytogenes cell wall as the pathogen transits from the extracellular environment to an intracytosolic lifestyle inside eukaryotic cells. Some of these changes include alterations in the relative amount and the mode of association of certain surface proteins.

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Section: Methodsmentioning

confidence: 99%

Section: Characterization Of Cell Wall Proteome Of L Monocytogenes Gmentioning

confidence: 99%

Association of ActA to Peptidoglycan Revealed by Cell Wall Proteomics of Intracellular Listeria monocytogenes

Portillo

Calvo

D'Orazio

et al. 2011

Journal of Biological Chemistry

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“…The sigB mutant derivative was obtained from Lone Brøndsted (Brondsted et al, 2003), and the perR mutant was constructed as described by Rea et al (2004). Strains were grown in rich growth medium (brain heart infusion, BHI; Oxoid) or improved minimal medium (Phan-Thanh & Gormon, 1997), with continuous shaking at 37 uC unless otherwise indicated. When required, erythromycin was added to a final concentration of 5 mg ml 21 .…”

Section: Methodsmentioning

confidence: 99%

The Dps-like protein Fri of Listeria monocytogenes promotes stress tolerance and intracellular multiplication in macrophage-like cells

Olsen¹,

Larsen²,

Gahan

et al. 2005

Microbiology

103

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Members of the ferritin-like Dps protein family are found in a number of bacterial species, where they demonstrate the potential to bind iron, and have been implicated in tolerance to oxidative stress. In this study of the food-borne pathogen Listeria monocytogenes, the fri gene encoding a Dps homologue was deleted, and, compared to wild-type cells, it was found that the resulting mutant was less resistant to hydrogen peroxide, and demonstrated reduced survival following long-term (7–11 days) incubation in laboratory media. In view of this, it is shown that fri gene expression is controlled by the hydrogen peroxide regulator PerR, as well as the general stress sigma factor σ B. When fri mutant cells were transferred to iron-limiting conditions, growth was retarded relative to wild-type cells, indicating that Fri may be required for iron storage. This notion is supported by the observation that the L. monocytogenes genome appears not to encode other ferritin-like proteins. Given the role of Fri in resistance to oxidative stress, and growth under iron-limiting conditions, the ability of the fri mutant to infect mice was examined. When injected by the intraperitoneal route, the fri mutant demonstrated a reduced capacity to proliferate in the organs of infected mice relative to the wild-type, whereas when the bacteria were supplied intravenously this effect was mitigated. In addition, the mutant was impaired in its ability to survive and grow in J774.A1 mouse macrophage cells. Thus, the data suggest that Fri contributes to the ability of L. monocytogenes to survive in environments where oxidative stress and low iron availability may impede bacterial proliferation.

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“…The bacteria from stock agar were first inoculated and grown at 37°C with agitation in BHI broth (DIFCO/OSI, Maurepas, France) for several generations (7-8 h), allowing the bacteria to recover from eventual damage or stress before being transferred to a synthetic medium for overnight culture. The synthetic medium used was specially designed for the optimal growth of Listeria and described elsewhere (Phan-Thanh and Gormon, 1997). An inoculum from BHI growth was diluted into 100 ml of synthetic medium to have about 10 5 cells per ml.…”

Section: Methodsmentioning

confidence: 99%

Physiological and biochemical aspects of the acid survival of Listeria monocytogenes.

Phan‐Thanh

Montagne

1998

J. Gen. Appl. Microbiol.

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A chemically defined minimal medium for the optimal culture of Listeria

Cited by 85 publications

References 10 publications

Association of ActA to Peptidoglycan Revealed by Cell Wall Proteomics of Intracellular Listeria monocytogenes

Association of ActA to Peptidoglycan Revealed by Cell Wall Proteomics of Intracellular Listeria monocytogenes

The Dps-like protein Fri of Listeria monocytogenes promotes stress tolerance and intracellular multiplication in macrophage-like cells

Physiological and biochemical aspects of the acid survival of Listeria monocytogenes.

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