The Legionella pneumophila F-box protein Lpp2082 (AnkB) modulates ubiquitination of the host protein parvin B and promotes intracellular replication

Lomma, Mariella; Dervins-Ravault, Delphine; Rolando, Monica; Nora, Tamara; Newton, Hayley J; Sansom, Fiona M.; Sahr, Tobias; Gómez-Valero, Laura; Jules, Matthieu; Hartland, Elizabeth L.; Buchrieser, Carmen

doi:10.1111/j.1462-5822.2010.01467.x

Cited by 143 publications

(211 citation statements)

References 92 publications

Supporting

Mentioning

211

Contrasting

Order By: Relevance

“…To validate these results, we assessed the effect of T2S on IL-6 production by the human macrophage-like cell line THP-1 as well as peripheral blood mononuclear cells (PBMCs) obtained from human volunteers. As is the case for U937 cells, THP-1 cells and PBMCs are widely utilized for studying the interaction between L. pneumophila and macrophages (27)(28)(29)(30)(31). For both of these host cells, infection with an lspF mutant that lacks T2S resulted in levels of IL-6 that were significantly higher than those obtained following infection with WT strain 130b ( Fig.…”

Section: Resultsmentioning

confidence: 88%

The Type II Secretion System of Legionella pneumophila Dampens the MyD88 and Toll-Like Receptor 2 Signaling Pathway in Infected Human Macrophages

2017

View full text Add to dashboard Cite

Previously, we reported that mutants of Legionella pneumophila lacking a type II secretion (T2S) system elicit higher levels of cytokines (e.g., interleukin-6 [IL-6]) following infection of U937 cells, a human macrophage-like cell line. We now show that this effect of T2S is also manifest upon infection of human THP-1 macrophages and peripheral blood monocytes but does not occur during infection of murine macrophages. Supporting the hypothesis that T2S acts to dampen the triggering of an innate immune response, we observed that the mitogen-activated protein kinase (MAPK) and nuclear transcription factor kappa B (NF-B) pathways are more highly stimulated upon infection with the T2S mutant than upon infection with the wild type. By using short hairpin RNA to deplete proteins involved in specific pathogen-associated molecular pattern (PAMP) recognition pathways, we determined that the dampening effect of the T2S system was not dependent on nucleotide binding oligomerization domain (NOD)-like receptors (NLRs), retinoic acid-inducible protein I (RIG-I)-like receptors (RLRs), double-stranded RNA (dsRNA)-dependent protein kinase receptor (PKR), or TIR domain-containing adaptor inducing interferon beta (TRIF) signaling or an apoptosis-associated speck-like protein containing a CARD (ASC)-or caspase-4-dependent inflammasome. However, the dampening effect of T2S on IL-6 production was significantly reduced upon gene knockdown of myeloid differentiation primary response 88 (MyD88), TANK binding kinase 1 (TBK1), or Toll-like receptor 2 (TLR2). These data indicate that the L. pneumophila T2S system dampens the signaling of the TLR2 pathway in infected human macrophages. We also document the importance of PKR, TRIF, and TBK1 in cytokine secretion during L. pneumophila infection of macrophages.KEYWORDS Legionella pneumophila, TLR2, cytokine, innate immunity, macrophage, type II secretion L egionella pneumophila, a Gram-negative bacterium that is widespread in aquatic habitats, is the principal agent of Legionnaires' disease pneumonia (1-6). In the lungs, Legionella bacteria invade and grow in resident macrophages and then trigger severe inflammation (2). In macrophages, L. pneumophila evades the degradative lysosomal pathway and replicates to large numbers within a membrane-bound vacuole, the Legionella-containing vacuole (LCV) (7,8). Two protein secretion systems, Lsp type II secretion (T2S) and Dot/Icm type IV secretion (T4S), play major roles in the pathogenesis of L. pneumophila (9, 10). In T2S, protein substrates are first translocated across the inner membrane, and upon the action of the T2S pilus-like apparatus, they then exit the bacterial cell through a specific outer membrane pore (11). Using proteomics and enzymatic assays, we have shown that the T2S system of L. pneumo-

show abstract

Section: Resultsmentioning

confidence: 88%

The Type II Secretion System of Legionella pneumophila Dampens the MyD88 and Toll-Like Receptor 2 Signaling Pathway in Infected Human Macrophages

2017

View full text Add to dashboard Cite

show abstract

“…Previously, several factors have been discovered that have subtle effects on bacterial replication in one or more models of infection 29,[48][49][50][51] . Although the larvae do not possess an adaptive immune system, the presence of the innate immune response provides stronger selection compared to macrophages alone, which may serve to amplify subtle phenotypes.…”

Section: Discussionmentioning

confidence: 99%

Use of <em>Galleria mellonella</em> as a Model Organism to Study <em>Legionella pneumophila</em> Infection

Harding

Schroeder

Collins

et al. 2013

JoVE

View full text Add to dashboard Cite

Citation: Harding, C.R., Schroeder, G.N., Collins, J.W., Frankel, G. Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection. J. Vis. Exp. (81), e50964, doi:10.3791/50964 (2013). AbstractLegionella pneumophila, the causative agent of a severe pneumonia named Legionnaires' disease, is an important human pathogen that infects and replicates within alveolar macrophages. Its virulence depends on the Dot/Icm type IV secretion system (T4SS), which is essential to establish a replication permissive vacuole known as the Legionella containing vacuole (LCV). L. pneumophila infection can be modeled in mice however most mouse strains are not permissive, leading to the search for novel infection models. We have recently shown that the larvae of the wax moth Galleria mellonella are suitable for investigation of L. pneumophila infection. G. mellonella is increasingly used as an infection model for human pathogens and a good correlation exists between virulence of several bacterial species in the insect and in mammalian models. A key component of the larvae's immune defenses are hemocytes, professional phagocytes, which take up and destroy invaders. L. pneumophila is able to infect, form a LCV and replicate within these cells. Here we demonstrate protocols for analyzing L. pneumophila virulence in the G. mellonella model, including how to grow infectious L. pneumophila, pretreat the larvae with inhibitors, infect the larvae and how to extract infected cells for quantification and immunofluorescence microscopy. We also describe how to quantify bacterial replication and fitness in competition assays. These approaches allow for the rapid screening of mutants to determine factors important in L. pneumophila virulence, describing a new tool to aid our understanding of this complex pathogen.

show abstract

“…One example is AnkB/LegAU13/Ceg27/Lpg2144/Lpp2082 that contains three different eukaryotic protein domains: an F-box domain, an ankyrin domain, and a CaaX motif ( Table 2). The F-box domain interacts with the SKP1 component of the SCF1 ubiquitin ligase complex and allows Legionella to exploit the host ubiquitination machinery, whereas the ankyrin motif, a widespread protein -protein interaction motif, probably interacts with the cellular target like Parvin B described for strain Paris (Price et al 2009;Lomma et al 2010). The F-box domain, as well as the two ankyrin protein-protein interaction domains, is essential for the biological function of AnkB.…”

Section: Eukaryotic-like Proteins Of Legionella Are Virulence Factorsmentioning

confidence: 99%

Genome Dynamics in Legionella: The Basis of Versatility and Adaptation to Intracellular Replication

Gómez-Valero

Buchrieser

2013

Cold Spring Harbor Perspectives in Medicine

Self Cite

View full text Add to dashboard Cite

The Legionella pneumophila F-box protein Lpp2082 (AnkB) modulates ubiquitination of the host protein parvin B and promotes intracellular replication

Abstract: SummaryThe environmental pathogen Legionella pneumophila encodes three proteins containing F-box domains and additional protein-protein interaction domains, reminiscent of eukaryotic SCF ubiquitin-

Cited by 143 publications

References 92 publications

The Type II Secretion System of Legionella pneumophila Dampens the MyD88 and Toll-Like Receptor 2 Signaling Pathway in Infected Human Macrophages

The Type II Secretion System of Legionella pneumophila Dampens the MyD88 and Toll-Like Receptor 2 Signaling Pathway in Infected Human Macrophages

Use of <em>Galleria mellonella</em> as a Model Organism to Study <em>Legionella pneumophila</em> Infection

Genome Dynamics in Legionella: The Basis of Versatility and Adaptation to Intracellular Replication

Contact Info

Product

Resources

About