Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans.
dc.contributor.author | Lemme, André | |
dc.contributor.author | Gröbe, Lothar | |
dc.contributor.author | Reck, Michael | |
dc.contributor.author | Tomasch, Jürgen | |
dc.contributor.author | Wagner-Döbler, Irene | |
dc.date.accessioned | 2011-11-25T14:53:36Z | |
dc.date.available | 2011-11-25T14:53:36Z | |
dc.date.issued | 2011-04 | |
dc.identifier.citation | Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans. 2011, 193 (8):1863-77 J. Bacteriol. | en |
dc.identifier.issn | 1098-5530 | |
dc.identifier.pmid | 21317319 | |
dc.identifier.doi | 10.1128/JB.01363-10 | |
dc.identifier.uri | http://hdl.handle.net/10033/190851 | |
dc.description.abstract | Competence-stimulating-peptide (CSP)-mediated competence development in Streptococcus mutans is a transient and biphasic process, since only a subpopulation induces the expression of ComX in the presence of CSP, and the activation of the DNA uptake machinery in this fraction shuts down ~3 to 4 h postinduction. Here, we combine for the first time, to our knowledge, the bacterial flow-cytometric sorting of cells and subpopulation-specific transcriptome analysis of both the competent and noncompetent fraction of CSP-treated S. mutans cells. Sorting was guided by a ComX-green fluorescent protein (ComX-GFP) reporter, and the transcriptome analysis demonstrated the successful combination of both methods, because a strong enrichment of transcripts for comX and its downstream genes was achieved. Three two-component systems were expressed in the competent fraction, and among them was ComDE. Moreover, the recently identified regulator system ComR/S was expressed exclusively in the competent fraction. In contrast, the expression of bacteriocin-related genes was at the same level in all cells. GFP reporter strains for ComE and CipB (mutacin V) confirmed this expression pattern on the single-cell level. Fluorescence microscopy revealed that some ComX-expressing cells committed autolysis in an early stage of competence initiation. In viable ComX-expressing cells, the uptake of DNA could be shown on the single-cell level. This study demonstrates that all cells in the population respond to CSP through the activation of bacteriocin-related genes. Some of these cells start to activate ComX expression but then segregate into two subpopulations, one becoming competent and another one that lyses, resulting in intrapopulation diversity. | |
dc.language.iso | en | en |
dc.subject.mesh | Artificial Gene Fusion | en |
dc.subject.mesh | Bacterial Proteins | en |
dc.subject.mesh | Flow Cytometry | en |
dc.subject.mesh | Gene Expression Profiling | en |
dc.subject.mesh | Gene Expression Regulation, Bacterial | en |
dc.subject.mesh | Genes, Reporter | en |
dc.subject.mesh | Green Fluorescent Proteins | en |
dc.subject.mesh | Streptococcus mutans | en |
dc.title | Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans. | en |
dc.type | Article | en |
dc.contributor.department | Helmholtz Centre for Infection Research, Research Group Microbial Communication, Braunschweig, Germany. Andre.Lemme@helmholtz-hzi.de | en |
dc.identifier.journal | Journal of bacteriology | en |
refterms.dateFOA | 2018-06-13T00:16:20Z | |
html.description.abstract | Competence-stimulating-peptide (CSP)-mediated competence development in Streptococcus mutans is a transient and biphasic process, since only a subpopulation induces the expression of ComX in the presence of CSP, and the activation of the DNA uptake machinery in this fraction shuts down ~3 to 4 h postinduction. Here, we combine for the first time, to our knowledge, the bacterial flow-cytometric sorting of cells and subpopulation-specific transcriptome analysis of both the competent and noncompetent fraction of CSP-treated S. mutans cells. Sorting was guided by a ComX-green fluorescent protein (ComX-GFP) reporter, and the transcriptome analysis demonstrated the successful combination of both methods, because a strong enrichment of transcripts for comX and its downstream genes was achieved. Three two-component systems were expressed in the competent fraction, and among them was ComDE. Moreover, the recently identified regulator system ComR/S was expressed exclusively in the competent fraction. In contrast, the expression of bacteriocin-related genes was at the same level in all cells. GFP reporter strains for ComE and CipB (mutacin V) confirmed this expression pattern on the single-cell level. Fluorescence microscopy revealed that some ComX-expressing cells committed autolysis in an early stage of competence initiation. In viable ComX-expressing cells, the uptake of DNA could be shown on the single-cell level. This study demonstrates that all cells in the population respond to CSP through the activation of bacteriocin-related genes. Some of these cells start to activate ComX expression but then segregate into two subpopulations, one becoming competent and another one that lyses, resulting in intrapopulation diversity. |