RNAi screen of
            <i>Salmonella</i>
            invasion shows role of COPI in membrane targeting of cholesterol and Cdc42

Misselwitz, Benjamin; Dilling, Sabrina; Vonaesch, Pascale; Sacher, Raphael; Snijder, Berend; Schlumberger, Markus C.; Samuel, Rout; Stark, Manuel; Mering, Christian von; Pelkmans, Lucas; Hardt, Wolf‐Dietrich

doi:10.1038/msb.2011.7

Cited by 84 publications

(110 citation statements)

References 97 publications

(159 reference statements)

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“…Using flow cytometry and confocal microscopy, we showed that Salmonella targeted mitotic cells more efficiently than cells in other phases of their cell cycle ( (Misselwitz et al, 2011). This was true for the different wild-type strains (12023, SL1344 and LT2) and cell lines tested (supplementary material Fig.…”

Section: Salmonella Invades Mitotic Cells Preferentiallymentioning

confidence: 89%

Preferential invasion of mitotic cells by Salmonella reveals that cell surface cholesterol is maximal during metaphase

Santos

Meinecke

Fessler

et al. 2013

Journal of Cell Science

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SummaryCell surface-exposed cholesterol is crucial for cell attachment and invasion of many viruses and bacteria, including the bacterium Salmonella, which causes typhoid fever and gastroenteritis. Using flow cytometry and 3D confocal fluorescence microscopy, we found that mitotic cells, although representing only 1-4% of an exponentially growing population, were much more efficiently targeted for invasion by Salmonella. This targeting was not dependent on the spherical shape of mitotic cells, but was instead SipB and cholesterol dependent. Thus, we measured the levels of plasma membrane and cell surface cholesterol throughout the cell cycle using, respectively, brief staining with filipin and a fluorescent ester of polyethylene glycol-cholesterol that cannot flip through the plasma membrane, and found that both were maximal during mitosis. This increase was due not only to the rise in global cell cholesterol levels along the cell cycle but also to a transient loss in cholesterol asymmetry at the plasma membrane during mitosis. We measured that cholesterol, but not phosphatidylserine, changed from a ,20:80 outer:inner leaflet repartition during interphase to ,50:50 during metaphase, suggesting this was specific to cholesterol and not due to a broad change of lipid asymmetry during metaphase. This explains the increase in outer surface levels that make dividing cells more susceptible to Salmonella invasion and perhaps to other viruses and bacteria entering cells in a cholesterol-dependent manner. The change in cholesterol partitioning also favoured the recruitment of activated ERM (Ezrin, Radixin, Moesin) proteins at the plasma membrane and thus supported mitotic cell rounding.

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Section: Salmonella Invades Mitotic Cells Preferentiallymentioning

confidence: 89%

Preferential invasion of mitotic cells by Salmonella reveals that cell surface cholesterol is maximal during metaphase

Santos

Meinecke

Fessler

et al. 2013

Journal of Cell Science

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“…For example, Shigella uses its IpaB effector protein to bind the host raft‐associated CD44 transmembrane receptor (Lafont et al , 2002); entry of Listeria monocytogenes into host cells requires the localization of the host receptors E‐cadherin and HGF‐R/Met in specific lipid domains (Seveau et al , 2004). In addition to receptors, plasma membrane composition itself, specifically cholesterol and sphingolipid membrane content, impacts the binding and internalization of various bacterial pathogens, including Shigella and Salmonella species (Garner et al , 2002; Lafont et al , 2002; Misselwitz et al , 2011a; Santos et al , 2013). …”

Section: Introductionmentioning

confidence: 99%

Stress‐induced host membrane remodeling protects from infection by non‐motile bacterial pathogens

et al. 2018

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While mucosal inflammation is a major source of stress during enteropathogen infection, it remains to be fully elucidated how the host benefits from this environment to clear the pathogen. Here, we show that host stress induced by different stimuli mimicking inflammatory conditions strongly reduces the binding of Shigella flexneri to epithelial cells. Mechanistically, stress activates acid sphingomyelinase leading to host membrane remodeling. Consequently, knockdown or pharmacological inhibition of the acid sphingomyelinase blunts the stress‐dependent inhibition of Shigella binding to host cells. Interestingly, stress caused by intracellular Shigella replication also results in remodeling of the host cell membrane, in vitro and in vivo, which precludes re‐infection by this and other non‐motile pathogens. In contrast, Salmonella Typhimurium overcomes the shortage of permissive entry sites by gathering effectively at the remaining platforms through its flagellar motility. Overall, our findings reveal host membrane remodeling as a novel stress‐responsive cell‐autonomous defense mechanism that protects epithelial cells from infection by non‐motile bacterial pathogens.

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“…On average, B2,400 cells were analysed per well and replicate. A Caenorhabditis elegans miRNA (cel-miR-231) with no sequence homology to any known human miRNA was used as the negative control in all experiments; a short interfering RNA (siRNA) against ACTR3, a protein important for the entry of Salmonella into host cells 17 , was used as the positive control ( Supplementary Fig. 1d).…”

Section: Screening For Mirnas Controlling Salmonella Infectionmentioning

confidence: 99%

“…The use of RNA interference screenings has improved our knowledge of the interplay between bacteria and host cells, identifying novel cellular factors critical for infection [17][18][19] . Only recently this approach has been extended to miRNAs 20,21 , and is yet to be applied for the identification of miRNAs critical for infection by bacterial pathogens.…”

mentioning

confidence: 99%

Functional high-throughput screening identifies the miR-15 microRNA family as cellular restriction factors for Salmonella infection

et al. 2014

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Increasing evidence suggests an important role for miRNAs in the molecular interplay\ud between bacterial pathogens and host cells. Here we perform a fluorescence microscopybased\ud screen using a library of miRNA mimics and demonstrate that miRNAs modulate\ud Salmonella infection. Several members of the miR-15 miRNA family were among the\ud 17 miRNAs that more efficiently inhibit Salmonella infection. We discovered that these\ud miRNAs are downregulated during Salmonella infection, through the inhibition of the\ud transcription factor E2F1. Analysis of miR-15 family targets revealed that derepression of\ud cyclin D1 and the consequent promotion of G1/S transition are crucial for Salmonella\ud intracellular proliferation. In addition, Salmonella induces G2/M cell cycle arrest in infected\ud cells, further promoting its replication. Overall, these findings uncover a mechanism whereby\ud Salmonella renders host cells more susceptible to infection by controlling cell cycle\ud progression through the active modulation of host cell miRNAs

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RNAi screen of Salmonella invasion shows role of COPI in membrane targeting of cholesterol and Cdc42

Abstract: A genome wide RNAi screen identifies 72 host cell genes affecting S. Typhimurium entry, including actin regulators and COPI. This study implicates COPI-dependent cholesterol and sphingolipid localization as a common mechanism of infection by bacterial and viral pathogens.

Cited by 84 publications

References 97 publications

Preferential invasion of mitotic cells by Salmonella reveals that cell surface cholesterol is maximal during metaphase

Preferential invasion of mitotic cells by Salmonella reveals that cell surface cholesterol is maximal during metaphase

Stress‐induced host membrane remodeling protects from infection by non‐motile bacterial pathogens

Functional high-throughput screening identifies the miR-15 microRNA family as cellular restriction factors for Salmonella infection

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