Cell differentiation defines acute and chronic infection cell types in Staphylococcus aureus
- University of Würzburg, Germany
- Newcastle University, United Kingdom
- Technical University of Munich, Germany
- University Hospital Würzburg, Germany
- National Center for Biotechnology, Spain
Figures
Figure 1 with 1 supplement
Staphylococcus aureus aggregates contain specialized cell types.
(A) Scheme of the agr signaling cascade in S. aureus. AIP binds to the AgrC histidine kinase membrane receptor and activates its cognate regulator AgrA via phosphorylation (AgrA~P). AgrA~P …
Figure 1—figure supplement 1
Staphylococcus aureus multicellular aggregates contain distinct cell types.
(A) Fluorescence microscopy image fields of S. aureus cells harboring transcriptional fusions of biofilm-associated ica/spa genes and planktonic psmα/β agr-related genes and, the transcriptional …
Figure 2 with 1 supplement
The agr QS system is an autonomous program for cell fate decision-making.
(A) Scheme showing the orthogonal system generated in Bacillus subtilis. In this system, the membrane kinase AgrC and its cognate regulator AgrA were expressed under the control of their own P2 …
Figure 2—figure supplement 1
Mathematical simulations of the agr orthologous system.
(A) Flow cytometry profiles of the orthogonal agr system in B. subtilis ΔsigB and control experiments with B. subtilis wild type strain and, when no AIP was added to the cultures. In this control …
Figure 3 with 1 supplement
Extracellular Mg2+increases cell wall rigidity in S. aureus.
(A) qRT-PCR assay to monitor σB activation using relative asp23 expression as readout (expression of this gene is dependent on σB activity) in TSB and TSBMg cultures of S. aureus strains of …
Figure 3—figure supplement 1
Extracellular Mg2+ activates σB stress regulon in S. auresus.
(A) Quantification of staphyloxantin production in TSBMg and TSB media was used as a proxy to monitor activation of σB, since the expression of the operon responsible for staphyloxanthin production …
Figure 4
AIP and Mg2+modulate the BRcell:DRcell ratio in S. aureus communities.
(A) Quantitative analysis of fluorescence microscopy images of agr-related promoters (Ppsmα and Ppsmβ) in TSBMg and TSB. We counted ~700 random cells from each of three independent microscopic …
Figure 5 with 2 supplements
Collective behavior of BRcells and DRcells in vitro.
(A) Mathematical modeling of WT and a matrix-deficient mutant (ΔicaΔspa) S. aureus growth in multicellular aggregates. Local concentrations of factors that affect AIP activity are represented on a …
Figure 5—figure supplement 1
Staphylococcus aureus develops architecturally complex multicellular aggregates in Magnesium-supplemented TSB medium (TSBMg).
(A) Local concentration of AIP and nutrients control the spatial distribution of replicative and the non-replicative cells in multicellular aggregates of S. aureus. Mathematical modeling of S. aureus…
Figure 5—figure supplement 2
Spatial distribution of BRcells and DRcells in vitro.
(A) Spatial localization of the distinct cell types within the morphologically different regions of the multicellular aggregates. Flow cytometry quantification of the size of each cell type of …
Figure 6 with 2 supplements
BRcell and DRcell subpopulations have different gene expression profiles.
(A) Unsupervised hierarchical clustering of commonly expressed genes differentially regulated in at least one of the libraries shows a specific, divergent expression profile for BRcells and DRcells. …
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Figure 6—source data 1
Genome-wide analysis of DRcell and BRcell sorted subpopulations.
Library specific variations are 0, showing that no statistically significant genetic differences were found between BRcells and DRcells genomes.
- https://doi.org/10.7554/eLife.28023.016
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Figure 6—source data 2
Gene quantification and differential expression analysis of BRcells and DRcells.
This data table contains the table for gene-wise quantification calculated for different cDNA libraries used in this study. DESeq analysis data for library comparisons.
- https://doi.org/10.7554/eLife.28023.017
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Figure 6—source data 3
Functional classification of annotated genes.
Gene Ontology classification of annotated genes that showed significant log2fold change value for differential expression generated by DESeq analysis. Aureowiki functional classification of the annotated genes that showed significant log2fold change value for differential expression generated by DESeq comparison analysis.
- https://doi.org/10.7554/eLife.28023.018
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Figure 6—source data 4
Hypergeometric analysis for library comparison comprising the log2fold values for differentially expressed genes that were shown to be expressed by DESeq analysis comparison of DRcells+ against BRcells+ sample sets.
Hypergeometric analysis probability and hypergeometric analysis heatmap comparison.
- https://doi.org/10.7554/eLife.28023.019
Figure 6—figure supplement 1
Experimental workflow to sort BRcells and DRcells using Fluorescence Activated Cell Sorting (FACS) to analyze and compare their transcriptomic profile.
(A) Schematic flow of the experimental approach used for cell sorting and RNA-sequencing of specific cell types. Multicellular aggregates were resuspended and disaggregated in RNAlater for cell …
Figure 6—figure supplement 2
Read alignment statistics, transcriptomic profile of BRcells and DRcells and differential distribution of cell types in distinct infected organs.
(A) Basic statistics of the samples. Number of reads obtained from sequencing; number of reads that passed the bioinformatics standard quality check and thus were aligned against the reference …
Figure 7 with 3 supplements
BRcells are more represented in infected kidneys and DRcells are more represented in infected hearts.
Histological preparations of infected kidneys (A) or hearts (B) stained with Giemsa solution and visualized using light microscopy. Upper row, 40X magnification of the preparation. Bar = 50 μm. The …
Figure 7—figure supplement 1
Bacterial loads in Mg2+-enriched and Mg2+-depleted organs.
(A) Bacterial loads of mice infected with and unlabeled S. aureus strain, Pica-yfp (BRcells) and Ppsmα-yfp (DRcells) labeled strains. (B) Upper panels, histological preparations of infected kidneys …
Figure 7—figure supplement 2
BRcells are more represented in infected kidneys and DRcells are more represented in infected hearts.
(A) Confocal microscopy images of infected kidneys and infected hearts, which were used for the quantitative analysis of Figure 7C,D. Left panels; bright field 63X-magnification images of different …
Figure 7—figure supplement 3
BRcells are more represented in infected kidneys and DRcells are more represented in infected hearts.
(A) Low magnification (10X) histological preparations of the infected kidneys (upper row) or hearts (lower panels) presented in Figure 7. Bar = 500 μm. The area delineated in red corresponds to the …
Figure 8
Low- and high-tagB strains display different infection patterns.
(A) Bacterial loads on different genetic backgrounds in kidney, heart and liver of infected mice. (B, C) qRT-PCR analysis of BRcell- (B) and DRcell- (C) related genes in kidney, heart and liver of …
Additional files
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Supplementary file 1
List of strains used in this study.
- https://doi.org/10.7554/eLife.28023.025
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Supplementary file 2
List of plasmids and plasmids used in this study.
- https://doi.org/10.7554/eLife.28023.026
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Transparent reporting form
- https://doi.org/10.7554/eLife.28023.027
