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dc.contributor.authorLemme, André
dc.contributor.authorGröbe, Lothar
dc.contributor.authorReck, Michael
dc.contributor.authorTomasch, Jürgen
dc.contributor.authorWagner-Döbler, Irene
dc.date.accessioned2011-11-25T14:53:36Z
dc.date.available2011-11-25T14:53:36Z
dc.date.issued2011-04
dc.identifier.citationSubpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans. 2011, 193 (8):1863-77 J. Bacteriol.en
dc.identifier.issn1098-5530
dc.identifier.pmid21317319
dc.identifier.doi10.1128/JB.01363-10
dc.identifier.urihttp://hdl.handle.net/10033/190851
dc.description.abstractCompetence-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.isoenen
dc.subject.meshArtificial Gene Fusionen
dc.subject.meshBacterial Proteinsen
dc.subject.meshFlow Cytometryen
dc.subject.meshGene Expression Profilingen
dc.subject.meshGene Expression Regulation, Bacterialen
dc.subject.meshGenes, Reporteren
dc.subject.meshGreen Fluorescent Proteinsen
dc.subject.meshStreptococcus mutansen
dc.titleSubpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research, Research Group Microbial Communication, Braunschweig, Germany. Andre.Lemme@helmholtz-hzi.deen
dc.identifier.journalJournal of bacteriologyen
refterms.dateFOA2018-06-13T00:16:20Z
html.description.abstractCompetence-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.


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