c-Myc plays a key role in IFN-γ-induced persistence of Chlamydia trachomatis
- Department of Microbiology, Biocenter, University of Wuerzburg, Germany
- German Cancer Research Center (DKFZ), Germany
- College of Veterinary Medicine, China Agricultural University, China
- Memorial Sloan Kettering Cancer Center, United States
- Department of Biochemistry and Molecular Biology, University of Wuerzburg, Germany
- Pathogen Biology, Rajiv Gandhi Centre for Biotechnology (RGCB), India
Figures
Figure 1 with 1 supplement
Interferon-gamma (IFN-γ) induces depletion of c-Myc and impairs chlamydial growth.
(A) HeLa 229 cells with an anhydrotetracycline (AHT)-inducible expression of shc-Myc were infected with Chlamydia trachomatis (Ctr) at multiplicity of infection (MOI) 1. The infected cells were …
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Figure 1—source data 1
Complete and cutted membranes of all Western blots from Figure 1.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig1-data1-v1.pdf
Figure 1—figure supplement 1
Interferon-gamma (IFN-γ) induces c-Myc depletion and impairs chlamydial growth.
(A) HeLa 229 NTC SCC5 cells with an anhydrotetracycline (AHT)-inducible expression of a control short hairpin RNA (shRNA) were infected with Chlamydia trachomatis (Ctr) at a multiplicity of …
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Figure 1—figure supplement 1—source data 1
Complete and cutted membranes of all Western blots from Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig1-figsupp1-data1-v1.pdf
Figure 2 with 1 supplement
Interferon-gamma (IFN-γ) induces depletion of c-Myc and impairs chlamydial growth in human fallopian tube organoids.
(A) Layout of the infectivity assay performed in human fallopian tube organoids. Organoids (see Materials and methods) were infected with Chlamydia trachomatis (Ctr) and treated with or without …
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Figure 2—source data 1
Complete and cutted membranes of all Western blots from Figure 2.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig2-data1-v1.pdf
Figure 2—figure supplement 1
Interferon-gamma (IFN-γ)-induced persistence of Chlamydia trachomatis (Ctr) in human organoids.
(A) Human fallopian tube organoids were infected with Ctr and treated with or without IFN-γ for 6 days. The infected organoids were lysed using glass beads, and dilutions of the supernatant were …
Figure 3 with 1 supplement
Expression of c-Myc rescues Chlamydia from persistence.
(A) WII-U2OS cells were induced with 100 ng/mL anhydrotetracycline (AHT) for 2 hr. Cells were left untreated or were pre-treated for 2 hr with 10 ng/mL interferon-gamma (IFN-γ), infected with Chlamyd…
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Figure 3—source data 1
Complete and cutted membranes of all Western blots from Figure 3.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig3-data1-v1.pdf
Figure 3—figure supplement 1
Expression of c-Myc rescues Chlamydia from persistence.
(A) For the same culture conditions as in Figure 3A, an infectivity assay was performed. The cells were lysed at 48 hpi and dilutions of the resulting Chlamydia-containing supernatant were added …
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Figure 3—figure supplement 1—source data 1
Complete and cutted membranes of all Western blots from Figure 3—figure supplement 1.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig3-figsupp1-data1-v1.pdf
Figure 4 with 1 supplement
L-tryptophan activates the pGSK3β-c-Myc axis and rescues chlamydial infection.
(A) HeLa 229 cells were treated with interferon-gamma (IFN-γ) and different doses of L-tryptophan (W) (10–100 ng/mL). After 30 hr the cells were lysed and used for Western blot analysis (n=3). (B) …
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Figure 4—source data 1
Complete and cutted membranes of all Western blots from Figure 4.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig4-data1-v1.pdf
Figure 4—figure supplement 1
L-tryptophan activates the pGSK3β-c-Myc axis and rescues chlamydial infection.
(A) HeLa 229 cells were either left uninfected or infected with Chlamydia (Ctr) and treated with interferon-gamma (IFN-γ) with or without L-tryptophan (W). After 30 hpi, the cells were analysed via …
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Figure 4—figure supplement 1—source data 1
Complete and cutted membranes of all Western blots from Figure 4—figure supplement 1.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig4-figsupp1-data1-v1.pdf
Figure 5 with 1 supplement
Influence of stabilized c-Myc on L-tryptophan (Trp) uptake and metabolites.
(A) HeLa 229 cells were infected with Chlamydia (Ctr) for various time points. The cells were analysed via Western blotting for the levels of LAT1 (n=3). (B) HeLa 229 cells were either left …
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Figure 5—source data 1
Complete and cutted membranes of all Western blots from Figure 5.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig5-data1-v1.pdf
Figure 5—figure supplement 1
Pathway analysis.
(A.–D) WII-U2OS cells were either left uninfected or infected with Chlamydia trachomatis (Ctr) at multiplicity of infection (MOI) 1 for 30 hr. The infected cells were either left untreated or …
Figure 6 with 1 supplement
Tricarboxylic acid (TCA) intermediates and nucleosides can overcome interferon-gamma (IFN-γ)-induced persistence.
(A) A heatmap with hierarchical clustering of all metabolites detected by LC-MS analysis of (Figure 5G). (B/C) WII-U2OS cells were either left uninfected or infected with Chlamydia trachomatis (Ctr) …
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Figure 6—source data 1
Complete and cutted membranes of all Western blots from Figure 6.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig6-data1-v1.pdf
Figure 6—figure supplement 1
Urea cycle intermediates.
(A) Cartoon depicting tricarboxylic acid (TCA) cycle and nucleotide biosynthesis in human cells. (B) Cartoon depicting the flux of aspartate from the TCA cycle. Oxaloacetate from TCA is converted …
Figure 7
The chlamydial Trp synthase operon is not involved in the rescue from persistence by c-Myc expression and tricarboxylic acid (TCA) metabolite supply.
(A) Western blot of wildtype Chlamydia trachomatis (Ctr) and TrpBA mutant to demonstrate the loss of TrpA and TrpB. (B) HeLa 229 cells were infected with Ctr or TrpBA mutant at multiplicity of …
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Figure 7—source data 1
Complete and cutted membranes of all Western blots from Figure 7.
- https://cdn.elifesciences.org/articles/76721/elife-76721-fig7-data1-v1.pdf
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/76721/elife-76721-transrepform1-v1.pdf
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Source data 1
Inclusion Forming Units (IFU).
- https://cdn.elifesciences.org/articles/76721/elife-76721-data1-v1.xlsx
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Source data 2
Metabolic profiling.
- https://cdn.elifesciences.org/articles/76721/elife-76721-data2-v1.xlsx
