Reverting the mode of action of the mitochondrial FOF1-ATPase by Legionella pneumophila preserves its replication niche
- Institut Pasteur, Biologie des Bactéries Intracellulaires and CNRS UMR 3525, France
- Faculté des Sciences, Université de Montpellier, France
- Faculté des Sciences, Université de Paris, France
- Sorbonne Université, Collège doctoral, France
Figures
Figure 1 with 1 supplement
Despite a reduction of oxidative phosphorylation (OXPHOS), human monocyte-derived macrophages (hMDMs) maintain their Δψm during infection by L. pneumophila.
(A) hMDMs were infected with L. pneumophila strain Paris (Lpp) wild-type (WT), a type 4 secretion system (T4SS)-deficient ΔdotA mutant, or left uninfected (noninfected). At 6 hr post-infection …
Figure 1—figure supplement 1
Oxidative phosphorylation (OXPHOS) and Δψm during infection by L. pneumophila.
(A) Bioenergetic profiles of the key parameters of mitochondrial respiration during a mitochondrial respiratory control assay using the Seahorse XF Mitostress kit. Sequential compound injections …
Figure 2 with 1 supplement
The mitochondrial FOF1-ATPase works in the ‘reverse mode’ during infection of human monocyte-derived macrophages (hMDMs) by L. pneumophila.
(A) In the ‘forward mode’ of the mitochondrial ATPase, the Δψm generated by the electron transport chain is used by the FOF1-ATPase to synthesize ATP. The ‘reverse mode’ of the FOF1-ATPase leads to …
Figure 2—figure supplement 1
Basal respiration of human monocyte-derived macrophages (hMDMs) infected with L. pneumophila Δspl mutant and validation of the model in HEK-293 cells.
(A) hMDMs were infected with L. pneumophila strain Paris WT (Lpp-WT), a type 4 secretion system (T4SS)-deficient ΔdotA mutant, a LpSpl-deficient Δspl mutant, or left uninfected (noninfected). At 6 …
Figure 3
Inhibition of the ‘reverse mode’ of mitochondrial FOF1 ATPase reduces the Δψm of L. pneumophila-infected human monocyte-derived macrophages (hMDMs).
(A) hMDMs were infected with GFP-expressing bacteria (green), Lpp-WT or Lpp-ΔdotA, or left uninfected (noninfected). At 5.5 hr post-infection (hpi), cells were labeled with Hoechst to identify cell …
Figure 4 with 2 supplements
Inhibition of FO-F1 ATPase ‘reverse mode’ increases cell death in L. pneumophila-infected human monocyte-derived macrophages (hMDMs).
(A) hMDMs were infected with Lpp-WT-GFP and were nontreated or treated with 50 μM BTB. The presence of GFP-expressing bacteria in each cell was monitored, and the number of living infected cells in …
Figure 4—figure supplement 1
Inhibition of FO-F1 ATPase ‘reverse mode’ delays cell death in L. pneumophila-infected human monocyte-derived macrophages (hMDMs), while transfection of LpSpl into HEK-293 cells protected transfected cells from Staurosporine (STS)-induced cell death.
(A) Noninfected hMDMs were stained with Hoechst to identify their nuclei and challenged with FCCP (10 μM) for 18 hr, while Annexin-V Alexa Fluor 647 was added to the cell culture to monitor cell …
Figure 4—figure supplement 2
BTB treatment, Δψm, and cell death of L. pneumophila-infected human monocyte-derived macrophages (hMDMs).
(A) hMDMs were infected with Lpp-WT-GFP (green), nuclei of host cells were stained with Hoechst (Nuc, blue), and tetramethylrhodamine methyl ester (TMRM) (red) and Annexin-V Alexa Fluor 647 (yellow) …
