Legionella dumoffii Utilizes Exogenous Choline for Phosphatidylcholine Synthesis

Palusińska-Szysz, Marta; Szuster‐Ciesielska, Agnieszka; Kania, Magdalena; Janczarek, Monika; Chmiel, Elżbieta; Danikiewicz, Witold

doi:10.3390/ijms15058256

Cited by 7 publications

(11 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, in contrast to L. pneumophila , L. dumoffii cultured on the choline-supplemented medium after treatment with apoLp-III showed a three times lower survival rate than the bacteria grown without choline supplementation [ 33 , 35 ]. We also reported on utilization of exogenous choline by L. dumoffii for synthesis of PC that was subsequently built into the outer and inner cell membrane [ 19 ]. A detailed lipidomic analysis revealed increased content of PC (and an accompanying decrease in PE) in cell membranes of L. dumoffii grown in the presence of choline as compared with non-supplemented bacteria [ 37 ].…”

Section: Discussionmentioning

confidence: 99%

“…Unlike L. pneumophila , L. dumoffii cells cultured in the presence of exogenous choline were considerably more susceptible, that is, they showed three times lower survival rate, after treatment with G. mellonella apoLp-III at a concentration of 0.4 mg/mL, than the bacteria grown without supplementation of this compound [ 33 ]. These results combined with the documented capability of utilization of exogenous choline for PC synthesis by Legionella [ 18 , 19 ] suggested that the increased PC content in the cell membrane could be responsible for the observed effects [ 33 ]. Indeed, a comparative lipidomic analysis revealed that cell membranes of L. dumoffii grown on choline-supplemented medium exhibited a ca.…”

Section: Introductionmentioning

confidence: 95%

“…It has been demonstrated that various Legionella spp. can synthesize PC from exogenous choline [ 17 , 18 , 19 , 20 ]. In L. dumoffii , structurally different PC species are localized in both the inner and outer membranes.…”

Section: Introductionmentioning

confidence: 99%

“…In L. dumoffii , structurally different PC species are localized in both the inner and outer membranes. Interestingly, L. dumoffii cultured in the presence of exogenous choline exhibits a reduced capability of TNF-α induction, which enables the bacteria to evade the host immune system [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Choline Supplementation Sensitizes Legionella dumoffii to Galleria mellonella Apolipophorin III

Palusińska-Szysz

Zdybicka-Barabas

Luchowski

et al. 2020

IJMS

View full text Add to dashboard Cite

The growth of Legionella dumoffii can be inhibited by Galleria mellonella apolipophorin III (apoLp-III) which is an insect homologue of human apolipoprotein E., and choline-cultured L. dumoffii cells are considerably more susceptible to apoLp-III than bacteria grown without choline supplementation. In the present study, the interactions of apoLp-III with intact L. dumoffii cells cultured without and with exogenous choline were analyzed to explain the basis of this difference. Fluorescently labeled apoLp-III (FITC-apoLp-III) bound more efficiently to choline-grown L. dumoffii, as revealed by laser scanning confocal microscopy. The cell envelope of these bacteria was penetrated more deeply by FITC-apoLp-III, as demonstrated by fluorescence lifetime imaging microscopy analyses. The increased susceptibility of the choline-cultured L. dumoffii to apoLp-III was also accompanied by alterations in the cell surface topography and nanomechanical properties. A detailed analysis of the interaction of apoLp-III with components of the L. dumoffii cells was carried out using both purified lipopolysaccharide (LPS) and liposomes composed of L. dumoffii phospholipids and LPS. A single micelle of L. dumoffii LPS was formed from 12 to 29 monomeric LPS molecules and one L. dumoffii LPS micelle bound two molecules of apoLp-III. ApoLp-III exhibited the strongest interactions with liposomes with incorporated LPS formed of phospholipids isolated from bacteria cultured on exogenous choline. These results indicated that the differences in the phospholipid content in the cell membrane, especially PC, and LPS affected the interactions of apoLp-III with bacterial cells and suggested that these differences contributed to the increased susceptibility of the choline-cultured L. dumoffii to G. mellonella apoLp-III.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Choline Supplementation Sensitizes Legionella dumoffii to Galleria mellonella Apolipophorin III

Palusińska-Szysz

Zdybicka-Barabas

Luchowski

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…All media and antibiotics were purchased from Sigma-Aldrich (Steinheim, Germany). For the experiments, the THP-1 cells were differentiated into macrophage-like cells as described earlier (Palusinska-Szysz et al, 2014). Briefly, THP-1 cells seeded at a density of 4 × 10 5 cells/ml in RPMI 1640 supplemented with 10% FCS onto 24-well plastic plates (Nunc, Roskilde, Denmark) were differentiated via 3-day exposure to 50 ng/ml phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) (Daigneault et al, 2010).…”

Section: Thp-1 Cell Culturesmentioning

confidence: 99%

The Role of Legionella pneumophila Serogroup 1 Lipopolysaccharide in Host-Pathogen Interaction

et al. 2019

Self Cite

View full text Add to dashboard Cite

The Legionella pneumophila TF3/1 mutant of the Corby strain, which possesses a point mutation in the active site of the O-acetyltransferase, synthesized the polysaccharide chain with a reduced degree of substitution with O-acetyl groups. The mutant did not produce a high-molecular-weight lipopolysaccharide (LPS) fraction above 12 kDa. The disturbances in LPS synthesis have an effect on the composition of other macromolecules (lipids and proteins), as indicated by differences in the infrared absorption spectra between the L. pneumophila Corby strain and its TF3/1 mutant. The wild-type strain contained less N +-CH 3 and C-N groups as well as more CH 3 groups than the mutant. The fatty acid composition showed that the wild type strain synthesized more branched acyl residues (a15:0, i16:0, and a17:0), a less unsaturated acid (16:1), and a straight-chain acid (18:0) than the mutant. The mutant synthesized approximately twice more a long-chain fatty acid (20:0) than the wild type. The main differences in the phospholipids between both strains were found in the classes of phosphatidylcholines and phosphatidylglycerols (PG). Substantial differences in the cell surface topography of these bacteria and their nanomechanical properties were shown by atomic force microscopy (AFM). The wild type strain had no undulated surface and produced numerous vesicles. In the case of the mutant type, the vesicles were not numerous, but there were grooves on the cell surface. The average roughness of the cell surface of the mutant was approximately twofold higher than in the wild-type strain. In turn, the wild-type strain exhibited much better adhesive properties than the mutant. The kinetic study of the interaction between the L. pneumophila strains and Acanthamoeba castellanii monitored by Förster resonance energy transfer revealed a pronounced difference, i.e., almost instantaneous and highly efficient binding of the L. pneumophila Corby strain to the amoeba surface, followed by penetration into the

show abstract

PCR method for the rapid detection and discrimination of Legionella spp. based on the amplification of pcs, pmtA, and 16S rRNA genes

Janczarek

Palusińska-Szysz

2015

J Appl Genetics

Self Cite

View full text Add to dashboard Cite

Legionella bacteria are organisms of public health interest due to their ability to cause pneumonia (Legionnaires' disease) in susceptible humans and their ubiquitous presence in water supply systems. Rapid diagnosis of Legionnaires' disease allows the use of therapy specific for the disease. L. pneumophila serogroup 1 is the most common cause of infection acquired in community and hospital environments. The non-L. pneumophila infections are likely under-detected because of a lack of effective diagnosis. In this work, simplex and duplex PCR assays with the use of new molecular markers pcs and pmtA involved in phosphatidylcholine synthesis were specified for rapid and cost-efficient identification and distinguishing Legionella species. The sets of primers developed were found to be sensitive and specific for reliable detection of Legionella belonging to the eight most clinically relevant species. Among these, four primer sets I, II, VI, and VII used for duplex-PCRs proved to have the highest identification power and reliability in the detection of the bacteria. Application of this PCR-based method should improve detection of Legionella spp. in both clinical and environmental settings and facilitate molecular typing of these organisms.

show abstract

Legionella dumoffii Utilizes Exogenous Choline for Phosphatidylcholine Synthesis

Cited by 7 publications

References 46 publications

Choline Supplementation Sensitizes Legionella dumoffii to Galleria mellonella Apolipophorin III

Choline Supplementation Sensitizes Legionella dumoffii to Galleria mellonella Apolipophorin III

The Role of Legionella pneumophila Serogroup 1 Lipopolysaccharide in Host-Pathogen Interaction

PCR method for the rapid detection and discrimination of Legionella spp. based on the amplification of pcs, pmtA, and 16S rRNA genes

Contact Info

Product

Resources

About