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| Status |
Public on Sep 07, 2013 |
| Title |
Differential expression of Dictyostelium discoideum AX2 upon infection with wt L. pneumophila JR32 vs. the L. pneumophila delta DotA mutant 24h p.i. |
| Organism |
Dictyostelium discoideum |
| Experiment type |
Expression profiling by array
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| Summary |
Differential gene expression of Dictyostelium discoideum after infection with Legionella pneumophila was investigated using DNA microarrays. A detailed analysis of the 24 h time point post infection was performed in comparison to three controls, uninfected cells and co-incubation with Legionella hackeliae and L. pneumophila DeltadotA. One hundred and thirty-one differentially expressed D. discoideum genes were identified as common to all three experiments and are thought to be involved in the pathogenic response. Functional annotation of the differentially regulated genes revealed that apart from triggering a stress response Legionella apparently not only interferes with intracellular vesicle fusion and destination but also profoundly influences and exploits the metabolism of its host. The results provide the basis for a better understanding of the complex host-pathogen interactions and for further studies on the Dictyostelium response to Legionella infection.
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| Overall design |
The bacterial strains used in this study were L. pneumophila Philadelphia I JR32, L. pneumophila Philadelphia I JR32 LELA 3118 (dotA3118:Tn903 DLL LacZ) and L. hackeliae (ATCC 35250). The Legionella strains were grown on buffered charcoal yeast extract agar (BCYE) at 37°C with 5% CO2 atmosphere for 3 days. The D. discoideum wild-type strain AX2 was grown at 23°C in 75 cm2 cell-culture flasks with 10 ml HL5 medium. For infection, Dictyostelium cells were harvested, resuspended in a 1:1 solution of HL5 medium and Soerensen buffer. Fifteen millilitres of a 1×10e6 cells/ml suspension were seeded into a 75 square-cm cell culture flask and the amoebae were inoculated with 10e7 bacteria/ml. Three different pairs of infection were compared: 1. AX2 infected with L. pneumophila JR32 versus uninfected cells; 19 microarrays of seven independent infections; 2. AX2 infected with L. pneumophila JR32 versus AX2 infected with L. pneumophila JR32 delta DotA; 4 microarrays of two independent infections; 3. AX2 infected with L. pneumophila JR32 versus AX2 infected with L. hackeliae; 4 microarrays of two independent infections. 24h post infection the RNA was isolated from 1.5×10e7 Dictyostelium cells and microarray analysis was performed as described (Farbrother et al., 2006).
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| Contributor(s) |
Farbrother P, Konertz R, Eichinger L |
| Citation(s) |
16469056 |
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| Submission date |
Sep 03, 2013 |
| Last update date |
Sep 18, 2013 |
| Contact name |
Ludwig Eichinger |
| E-mail(s) |
ludwig.eichinger@uni-koeln.de
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| Phone |
+49 221 478 6928
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| Organization name |
Institute for Biochemistry 1
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| Lab |
AG Eichinger
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| Street address |
Joseph-Stelzmann-Strasse 52
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| City |
Cologne |
| ZIP/Postal code |
50931 |
| Country |
Germany |
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| Platforms (1) |
| GPL1972 |
Dictyostelium discoideum cDNA Microarray 6000 |
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| Samples (4)
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| GSM1220865 |
SAE 12650151 Prep. 1 Cy5 t24 vs. Prep. 3 Cy3 t24 |
| GSM1220866 |
SAE 12650152 Prep. 1 Cy3 t24 vs. Prep. 3 Cy5 t24 |
| GSM1220867 |
SAE 12650153 Prep. 2 Cy5 t24 vs. Prep. 4 Cy3 t24 |
| GSM1220868 |
SAE 12650154 Prep. 2 Cy3 t24 vs. Prep. 4 Cy5 t24 |
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| This SubSeries is part of SuperSeries: |
| GSE50680 |
Dictyostelium transcriptional host cell response upon infection with Legionella |
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| Relations |
| BioProject |
PRJNA218459 |
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