Dynamic subcellular localization of a respiratory complex controls bacterial respiration
- Institut de Microbiologie de la Méditerranée, Centre national de la recherche scientifique, Aix Marseille Université, France
Figures
Figure 1
The GFP-labeled nitrate reductase complex is active and fully assembled
.(A) Growth curves of E. coli strains expressing untagged or tagged-NarGHI under nitrate-respiring conditions. Cells were grown anaerobically in a minimal medium using glycerol as sole carbon source …
Figure 2 with 1 supplement
The GFP-labeled nitrate reductase complex concentrates at the cell poles under nitrate-respiring conditions.
(A) Fluorescence images (top) and overlays of fluorescence and phase contrast images (bottom) are shown for nitrate-respiring and oxygen-respiring cells. The deconvolved image of the fluorescence …
Figure 2—figure supplement 1
Spatial distribution of clusters.
(A) A density map was built from a two-dimensional histogram (cell length vs relative cluster position) of the fluorescence signal observed in nitrate-respiring or oxygen-respiring LCB3635 cells. …
Figure 3 with 1 supplement
Metabolism-dependent localization of the nitrate reductase OXPHOS complex.
Mean frequency of LCB3635 cells displaying clusters at the cell poles for various metabolic conditions (anaerobic respiration with either nitrate, fumarate or TMAO as terminal electron acceptor, …
Figure 3—figure supplement 1
Comparative analysis of cells displaying clusters at the cell poles vs cells displaying clusters along the cell body upon varying metabolic conditions.
(Gray bars) Mean frequency of LCB3635 cells displaying clusters of the fluorescence signal at the cell poles. (Black bars) Mean frequency of LCB3635 cells displaying clusters of the fluorescence …
Figure 4
Proton gradient is a cue for polar localization of the anaerobic OXPHOS complex.
(A) The pmf is important for polar localization of the nitrate reductase complex. Shown is the mean frequency of nitrate-respiring LCB3635 cells displaying clusters at the cell poles upon addition …
Figure 5 with 2 supplements
Activity of the nitrate reductase OXPHOS complex is not essential for polar positioning unless a ΔpH is established.
(A) Fluorescence images of JCB4023/pVA70GFP cells expressing active (top) or inactive NarG-H50S variant of the GFP-labeled nitrate reductase (bottom) are shown. Cells were grown anaerobically either …
Figure 5—figure supplement 1
Activity of the nitrate reductase OXPHOS complex is not essential for polar positioning unless a ΔpH is established.
Fluorescence images of JCB4023/pVA70GFP cells expressing inactive NarG-H50S variant of the GFP-labeled nitrate reductase and grown anaerobically either in glycerol-nitrate (nitrate) or …
Figure 5—figure supplement 2
Cells expressing the GFP-labeled nitrate reductase complex from the pVA70GFP plasmid show identical metabolism-dependent localization of the complex.
(A) Overlays of fluorescence and phase contrast images are shown for nitrate-respiring and oxygen-respiring JCB4023 cells expressing the GFP-tagged NarGHI complex from the pVA70GFP plasmid. (B) Mean …
Figure 6 with 1 supplement
Metabolism-dependent localization changes occur in a timeframe of several tens of minutes.
(A) (Black bars) Nitrate-respiring JCB4023/pVA70GFP cells were submitted to strong aeration and the fluorescence pattern was evaluated at 15-min intervals. (Gray bars) Oxygen-respiring …
Figure 6—figure supplement 1
Metabolism-dependent localization changes occur in a timeframe of several tens of minutes.
(A) Changes in GFP-labeled nitrate reductase content as estimated by immunoblotting. (Black bars) anoxic–oxic transition. (Gray bars) oxic–anoxic transition. (B) Changes in GFP-labeled nitrate …
Figure 7
Polar localization determines integration of the nitrate reductase complex in anaerobic respiratory chains.
(A) Constant activity of the GFP-tagged complex upon distinct subcellular localization. Quinol:nitrate oxidoreductase activity was measured on membranes prepared from oxygen-respiring or …
Additional files
-
Supplementary file 1
List of strains, plasmids, and oligonucleotides.
- https://doi.org/10.7554/eLife.05357.015
