Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans.

2.50
HDL Handle:
http://hdl.handle.net/10033/190851
Title:
Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans.
Authors:
Lemme, André; Gröbe, Lothar; Reck, Michael; Tomasch, Jürgen; Wagner-Döbler, Irene
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.
Affiliation:
Helmholtz Centre for Infection Research, Research Group Microbial Communication, Braunschweig, Germany. Andre.Lemme@helmholtz-hzi.de
Citation:
Subpopulation-specific transcriptome analysis of competence-stimulating-peptide-induced Streptococcus mutans. 2011, 193 (8):1863-77 J. Bacteriol.
Journal:
Journal of bacteriology
Issue Date:
Apr-2011
URI:
http://hdl.handle.net/10033/190851
DOI:
10.1128/JB.01363-10
PubMed ID:
21317319
Type:
Article
Language:
en
ISSN:
1098-5530
Appears in Collections:
Publications of RG Microbial Communication (KOM)

Full metadata record

DC FieldValueLanguage
dc.contributor.authorLemme, Andréen
dc.contributor.authorGröbe, Lotharen
dc.contributor.authorReck, Michaelen
dc.contributor.authorTomasch, Jürgenen
dc.contributor.authorWagner-Döbler, Ireneen
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.en
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

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