A frameshift in Yersinia pestis rcsD alters canonical Rcs signalling to preserve flea-mammal plague transmission cycles
- NHC key laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, China
- Department of Basic Medical Sciences, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, China
- Paul G. Allen School for Global Health, Washington State University, United States
- State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, China
Figures
Figure 1
rcsDpe negatively regulates biofilm formation, while rcsDpstb positively regulates biofilm formation in Y. pestis.
(A) Schematic representation of the rcsD frameshift mutation that occurred during speciation of Y. pestis from its ancestor Y. pseudotuberculosis. rcsDpstb: rcsD from Y. pseudotuberculosis; rcsDpe: r…
Figure 2 with 2 supplements
Expression of RcsD-Hpt is initiated by an uncommon AUU start codon and negatively regulates biofilm formation in Y. pestis.
(A) Schematic representation of mutations introduced to the putative RBS and start codon of RcsD-Hpt. ATT–312→GGT or ATT–312→ATG indicate that the predicted ATT start codon was mutated to GGT or …
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Figure 2—source data 1
Raw source data for Figure 2D.
- https://cdn.elifesciences.org/articles/83946/elife-83946-fig2-data1-v2.zip
Figure 2—figure supplement 1
Characterization of proteins encoded by rcsDpe region, related to Figure 2.
(A) Schematic representation of mutations introduced to the putative but non-functional start codons. (B) Crystal violet (CV) biofilm assay using wild type (WT) and ΔrcsD strains complemented by the …
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Figure 2—figure supplement 1—source data 1
Raw source data for Figure 2—figure supplement 1E.
The original file of the full raw unedited blots and the uncropped blot with the relevant bands clearly labelled as Figure 2—figure supplement 1E.
- https://cdn.elifesciences.org/articles/83946/elife-83946-fig2-figsupp1-data1-v2.zip
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Figure 2—figure supplement 1—source data 2
Raw source data for Figure 2—figure supplement 1F.
The original file of the full raw unedited blots and the uncropped blot with the relevant bands clearly labelled as Figure 2—figure supplement 1F.
- https://cdn.elifesciences.org/articles/83946/elife-83946-fig2-figsupp1-data2-v2.zip
Figure 2—figure supplement 2
The conserved His residue, H844, is important for the function of RcsD and RcsD-Hpt.
(A) Crystal violet (CV) assay of Y. pestis expressing rcsD and its H844A-relevant derivatives. CV assays in panel C of Figure 2—figure supplement 1 and panel A of Figure 2—figure supplement 2 were …
Figure 3
The frameshift mutation in rcsD alters the Rcs signalling pathway in Y. pestis.
Congo red (CR) pigmentation assay (A and C) and crystal violet (CV) biofilm assay (B and D) the derivatives of Y. pestis KIM6+ (wild type [WT]) and the rcsDpstb substitution strain. prcsF, plasmid …
Figure 4
The frameshift mutation in rcsD increases the phosphorylation of RcsB and represses the expression of HmsT.
(A) Crystal violet (CV) biofilm assays using a wild type (WT) (white), ΔrcsB (grey), or rcsB (D56Q) (black) mutant strain, each harbouring pUC19 vectors expressing rcsDpe or rcsDpstb. (B) …
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Figure 4—source data 1
Raw source data for Figure 4B.
The original file of the full raw unedited blots and the uncropped blot with the relevant bands clearly labelled as Figure 4B.
- https://cdn.elifesciences.org/articles/83946/elife-83946-fig4-data1-v2.zip
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Figure 4—source data 2
Raw source data for Figure 4D.
The original file of the full raw unedited blots and the uncropped blot with the relevant bands clearly labelled as Figure 4D.
- https://cdn.elifesciences.org/articles/83946/elife-83946-fig4-data2-v2.zip
Figure 5
The frameshift mutation in rcsD stabilizes the pgm locus in Y. pestis.
(A) Bacterial burdens in fleas infected with Y. pestis wild type (WT), rcsDpe::rcsDpstb and rcsDN-term strains after 0, 7, and 14 days of infection. (B) Cumulative blockage of fleas after 4 weeks of …
Figure 6 with 1 supplement
Genome-wide identification of genes regulated by the Rcs phosphorelay system in Y. pestis.
(A–C) Survival of C57BL/6 mice infected with Y. pestis Microtus strain 201 and its derivatives using an infectious dose of 40, 160, and 800 colony-forming units (CFU). (D) Venn diagram of …
Figure 6—figure supplement 1
RNA-seq analysis of genes regulated by Rcs system, related to Figure 6.
(A–D) Clusters of orthologous groups (COG) analysis of RNA-seq gene expression data comparing the ΔrcsB strain and wild type (WT) (A), and rcsDpe::rcsDpstb with WT (B) at 26°C; ΔrcsB with WT (C), …
Figure 7
Genetic changes in Rcs genes during speciation of Y. pestis.
Acquisition events are indicated by green lightning symbols, loss of genetic material by a red cross, and mutation by blue circles. All comparisons shown are relative to Y. pseudotuberculosis. …
Figure 8 with 1 supplement
Predicted model of Rcs signal transduction and phosphoryl transfer in Y. pseudotuberculosis and Y. pestis.
In Y. pseudotuberculosis, full-length RcsD plays a dominant role while the lowly expressed RcsD-Hpt might play a moonlighting function. When Rcs is in ‘off’ state, RcsF is not activated leading to …
Figure 8—figure supplement 1
Evolutionary model for how genetic changes in the Rcs system fine-tunes biofilm formation to preserve virulence of flea bite-transmitted Y. pestis.
Functional rcsA and intact rcsD are encoded by ancestral Y. pseudotuberculosis. Biofilm formation was strongly repressed by the Rcs system due to the presence of RcsA. Y. pseudotuberculosis was …
Additional files
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Supplementary file 1
Strains used in this study.
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp1-v2.docx
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Supplementary file 2
Plasmids used in this study.
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp2-v2.docx
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Supplementary file 3
Primers and oligos used in this study.
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp3-v2.docx
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Supplementary file 4
Differential expression of Y. pestis genes in Y. pestis mutant strains growing at temperatures (26°C or 37°C).
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp4-v2.xlsx
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Supplementary file 5
Mutations in Y. pestis during evolution involved in this study.
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp5-v2.docx
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Supplementary file 6
Putative SD and start codon sites in rcsD in Enterobacteriaceae.
- https://cdn.elifesciences.org/articles/83946/elife-83946-supp6-v2.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/83946/elife-83946-mdarchecklist1-v2.pdf
