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| Status |
Public on Sep 25, 2013 |
| Title |
Differential expression of Dictyostelium discoideum AX2 upon infection with wt L. pneumophila JR32 vs. uninfected 48h p.i. (timecourse experiment) |
| 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 in comparison to uninfected cells was investigated using DNA microarrays. Investigation of a 48 h time course of infection revealed several clusters of co-regulated genes, an enrichment of preferentially up- or downregulated genes in distinct functional categories and also showed that most of the transcriptional changes occurred 24 h after infection. 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. For some of the identified genes, e.g. rtoA involvement in the host response has been demonstrated in a recent study, for others such a role appears plausible. 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 strain used in this study was L. Pneumophila Philadelphia I JR32. The strain was 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 cme2 cell culture flask and the amoebae were inoculated with 10e7 bacteria/ml. After different time intervals of incubation (1, 3, 6, 24, and 48 h) the RNA was isolated from 1.5×10e7 Dictyostelium cells. Usually two or three parallel cultures for the experiment and the control were inoculated per infection. The percentage of infected cells was determined by in situ hybridization with Legionella-specific 16S rRNA probes. The average from three independent determinations was 34, 42, 42, 57 and 82% of infected Dictyostelium cells after 1, 3, 6, 24 and 48 h, respectively.
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| Contributor(s) |
Farbrother P, Konertz R, Eichinger L |
| Citation(s) |
16469056 |
| |
| Submission date |
Sep 18, 2013 |
| Last update date |
Sep 25, 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 (6)
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| GSM1233386 |
SAE 12523244 Prep. 2 L.p. JR32 Cy5 vs. Prep. 2 uninfected Cy3 t48 |
| GSM1233387 |
SAE 12523245 Prep. 3 L.p. JR32 Cy3 vs. Prep. 3 uninfected Cy5 t48 |
| GSM1233388 |
SAE 12523246 Prep. 3 L.p. JR32 Cy5 vs. Prep. 3 uninfected Cy3 t48 |
| GSM1233389 |
SAE 12523357 Prep. 1 L.p. JR32 Cy3 vs. Prep. 1 uninfected Cy5 t48 |
| GSM1233390 |
SAE 12522855 Prep. 2 L.p. JR32 Cy3 vs. Prep. 2 uninfected Cy5 t48 |
| GSM1233391 |
SAE 12523227 Prep. 1 L.p. JR32 Cy5 vs. Prep. 1 uninfected Cy3 t48 |
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| This SubSeries is part of SuperSeries: |
| GSE51185 |
Differential expression of Dictyostelium discoideum AX2 upon infection with wt L. pneumophila JR32 vs. uninfected (timecourse experiments) |
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| Relations |
| BioProject |
PRJNA221557 |
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