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A miRNA-mediated cellular network controls Legionella pneumophila replication in human macrophages via regulation of LGALS8 and MX1

ABSTRACT: Proteome profiles of THP-1 cells with or without pre-treatment with selected miRNA upon infection with L. pertussis were characterized and quantified using SILAC.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Kristin Surmann

LAB HEAD: Prof. Dr. Uwe Völker

PROVIDER: PXD012315 | Pride | 2020-09-28

REPOSITORIES: pride

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	160408_SB_KRIS_Capsys_452_1_F1_F2_24h_scr.raw	Raw
	160408_SB_KRIS_Capsys_452_1_F3_F4_24h_scr.raw	Raw
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	160408_SB_KRIS_Capsys_452_1_F7_F8_24h_scr.raw	Raw
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Publications

Interaction of Staphylococcus aureus and Host Cells upon Infection of Bronchial Epithelium during Different Stages of Regeneration.

Palma Medina Laura M LM Becker Ann-Kristin AK Michalik Stephan S Surmann Kristin K Hildebrandt Petra P Gesell Salazar Manuela M Mekonnen Solomon A SA Kaderali Lars L Völker Uwe U van Dijl Jan Maarten JM

ACS infectious diseases 20200708 8

The primary barrier that protects our lungs against infection by pathogens is a tightly sealed layer of epithelial cells. When the integrity of this barrier is disrupted as a consequence of chronic pulmonary diseases or viral insults, bacterial pathogens will gain access to underlying tissues. A major pathogen that can take advantage of such conditions is Staphylococcus aureus, thereby causing severe pneumonia. In this study, we investigated how S. aureus responds to different cond ...[more]

PMID: 32579327

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Translocated Legionella pneumophila small RNAs mimic eukaryotic miRNAs to dampen the host immune response [THP-1]

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Screening for eukaryotic motives in Legionella pneumophila reveals bacterial LPC_1677 as functional host histone de-acetylase

Project description:Purpose: The interaction between Legionella pneumophila and its host cell is not completly understood. The aim of this study was to identify eukaryotic-like bacterial factors which manipulate the host cell. Method: A stable overexpression of LPC_1677 was performed in THP-1 cells. The LPC_1677 overexpressing cells were infected with L.p. and RNA sequencing was performed. Key pro-inflammatory markers were validated via qPCR and ELISA. Results: Bioinformatic screening for eukaryotic-like motives in L.p. took us to LPC_1677 which deacetylates host histones and thereby attenuating gene transcription. Conclusion: This study demonstrated that the eukaryotic-like bacterial factor LPC_1677 de-acetylates Histones in the host cell.

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Use of partial 16S rRNA gene sequencing for identification of Legionella pneumophila and non-pneumophila Legionella spp.

Project description:We examined 49 Legionella species, 26 L. pneumophila and 23 non-pneumophila Legionella spp., using partial 16S rRNA gene sequencing. This approach accurately identified all the L. pneumophila isolates, characterized all non-pneumophila Legionella isolates as such within this genus, and classified most (20/23; 87%) of the non-pneumophila Legionella isolates to the species level.

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Legionellosis from Legionella pneumophila serogroup 13.

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A multi-center validation study on the discrimination of Legionella pneumophila sg.1, Legionella pneumophila sg. 2-15 and Legionella non-pneumophila isolates from water by FT-IR spectroscopy

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Legionella pneumophila proteome

Project description:Legionella pneumophila cells were harvested during exponential growth (RP) and stationary growth (TP). VBNC cells were also anylzed. Protein subfractions were studied.

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Legionella pneumophila transcriptomic response to CuO nanoparticles

Project description:Legionella pneumophila was exposed for 3h to 160 ug/ml of CuO nanoparticle. Untreated samples are used as the control condition.

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Transcriptomic change of Legionella pneumophila in water.

Project description:Legionella pneumophila (Lp) is an opportunistic pathogen and its survival in water is critical for human infection. Therefore, identifying the genes of Lp that are required for survival in water may help devise strategies to prevent Legionella outbreaks. In this study, we exposed Lp in rich medium and in an artificial freshwater medium (Fraquil) for 2, 6 and 24 hours to uncover the global transcriptomic changes of Lp in water. The repression of major metabolic pathways, such as division, transcription and translation, suggests that Lp enters a dormant state in water. The induction of the flagellar associated genes (flg, fli and mot), enhance entry genes (enh) and some Icm/Dot effectors suggests that Lp may be waiting to establish intracellular replication in suitable host. Moreover, many genes involved in resistance to antibiotic and oxidative stress were induced, suggesting that Lp may be more tolerant to environmental stresses in water. Indeed, Lp exposed to water is more resistant to erythromycin, gentamycin and kanamycin than those cultured in rich medium. Apart from this, the gene bdhA involved in the degradation of the intracellular energy storage compound poly-hydroxybutyrate is highly expressed in water. Further characterization shows that bdhA is positively regulated by RpoS during short-term exposure to water. The deletion mutant of bdhA had a survival defect in water at 37°C, demonstrating that this gene is important for maintaining the long-term survivorship of Lp in water. Other identified genes highly induced upon exposure to water could also be necessary for Lp to survive in water.

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Project description:Legionella pneumophila is a Gram-negative opportunistic human pathogen that infects and multiplies in a broad range of phagocytic protozoan and mammalian phagocytes. Based on the observation that small regulatory RNAs (sRNAs) play an important role in controlling virulence-related genes in several pathogenic bacteria, we attempted to identify sRNAs expressed by L. pneumophila. We used computational prediction followed by experimental verification to identify and characterize sRNAs encoded in the L. pneumophila genome. A 50-mer probe microarray was constructed to test the expression of predicted sRNAs in bacteria grown under a variety of conditions. This strategy successfully identified 22 expressed RNAs, out of which 6 were confirmed by northern blot and RACE. One of the identified sRNAs is highly expressed in postexponential phase, and computational prediction of its secondary structure reveals a striking similarity to the structure of 6S RNA, a widely distributed prokaryotic sRNA, known to regulate the activity of sigma(70)-containing RNA polymerase. A 70-mer probe microarray was used to identify genes affected by L. pneumophila 6S RNA in stationary phase. The 6S RNA positively regulates expression of genes encoding type IVB secretion system effectors, stress response genes such as groES and recA, as well as many genes involved in acquisition of nutrients and genes with unknown or hypothetical functions. Deletion of 6S RNA significantly reduced L. pneumophila intracellular multiplication in both protist and mammalian host cells, but had no detectable effect on growth in rich media.

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