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dc.contributor.authorBlanka, Andrea
dc.contributor.authorSchulz, Sebastian
dc.contributor.authorEckweiler, Denitsa
dc.contributor.authorFranke, Raimo
dc.contributor.authorBielecka, Agata
dc.contributor.authorNicolai, Tanja
dc.contributor.authorCasilag, Fiordiligie
dc.contributor.authorDüvel, Juliane
dc.contributor.authorAbraham, Wolf-Rainer
dc.contributor.authorKaever, Volkhard
dc.contributor.authorHäussler, Susanne
dc.date.accessioned2015-01-16T15:07:52Z
dc.date.available2015-01-16T15:07:52Z
dc.date.issued2014-01
dc.identifier.citationIdentification of the alternative sigma factor SigX regulon and its implications for Pseudomonas aeruginosa pathogenicity. 2014, 196 (2):345-56 J. Bacteriol.en
dc.identifier.issn1098-5530
dc.identifier.pmid24187091
dc.identifier.doi10.1128/JB.01034-13
dc.identifier.urihttp://hdl.handle.net/10033/338491
dc.description.abstractPseudomonas aeruginosa is distinguished by its broad metabolic diversity and its remarkable capability for adaptation, which relies on a large collection of transcriptional regulators and alternative sigma (σ) factors. The largest group of alternative σ factors is that of the extracytoplasmic function (ECF) σ factors, which control key transduction pathways for maintenance of envelope homeostasis in response to external stress and cell growth. In addition, there are specific roles of alternative σ factors in regulating the expression of virulence and virulence-associated genes. Here, we analyzed a deletion mutant of the ECF σ factor SigX and applied mRNA profiling to define the SigX-dependent regulon in P. aeruginosa in response to low-osmolarity-medium conditions. Furthermore, the combination of transcriptional data with chromatin immunoprecipitation (ChIP) followed by high-throughput sequencing (ChIP-seq) led to the identification of the DNA binding motif of SigX. Genome-wide mapping of SigX-binding regions revealed enrichment of downstream genes involved in fatty acid biosynthesis, type III secretion, swarming and cyclic di-GMP (c-di-GMP) signaling. In accordance, a sigX deletion mutant exhibited altered fatty acid composition of the cell membrane, reduced cytotoxicity, impaired swarming activity, elevated c-di-GMP levels, and increased biofilm formation. In conclusion, a combination of ChIP-seq with transcriptional profiling and bioinformatic approaches to define consensus DNA binding sequences proved to be effective for the elucidation of the regulon of the alternative σ factor SigX, revealing its role in complex virulence-associated phenotypes in P. aeruginosa.
dc.language.isoenen
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/260276/en
dc.rightsopenAccessen
dc.subject.meshBinding Sitesen
dc.subject.meshChromatin Immunoprecipitationen
dc.subject.meshCulture Mediaen
dc.subject.meshGene Deletionen
dc.subject.meshGene Expression Profilingen
dc.subject.meshGene Expression Regulation, Bacterialen
dc.subject.meshHigh-Throughput Nucleotide Sequencingen
dc.subject.meshOsmotic Pressureen
dc.subject.meshProtein Bindingen
dc.subject.meshPseudomonas aeruginosaen
dc.subject.meshRegulonen
dc.subject.meshSigma Factoren
dc.subject.meshVirulence Factorsen
dc.titleIdentification of the alternative sigma factor SigX regulon and its implications for Pseudomonas aeruginosa pathogenicity.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre of infection research; Inhoffenstr. 7; D38124 Braunschweig; Germany.en
dc.identifier.journalJournal of bacteriologyen
refterms.dateFOA2018-06-13T05:35:17Z
html.description.abstractPseudomonas aeruginosa is distinguished by its broad metabolic diversity and its remarkable capability for adaptation, which relies on a large collection of transcriptional regulators and alternative sigma (σ) factors. The largest group of alternative σ factors is that of the extracytoplasmic function (ECF) σ factors, which control key transduction pathways for maintenance of envelope homeostasis in response to external stress and cell growth. In addition, there are specific roles of alternative σ factors in regulating the expression of virulence and virulence-associated genes. Here, we analyzed a deletion mutant of the ECF σ factor SigX and applied mRNA profiling to define the SigX-dependent regulon in P. aeruginosa in response to low-osmolarity-medium conditions. Furthermore, the combination of transcriptional data with chromatin immunoprecipitation (ChIP) followed by high-throughput sequencing (ChIP-seq) led to the identification of the DNA binding motif of SigX. Genome-wide mapping of SigX-binding regions revealed enrichment of downstream genes involved in fatty acid biosynthesis, type III secretion, swarming and cyclic di-GMP (c-di-GMP) signaling. In accordance, a sigX deletion mutant exhibited altered fatty acid composition of the cell membrane, reduced cytotoxicity, impaired swarming activity, elevated c-di-GMP levels, and increased biofilm formation. In conclusion, a combination of ChIP-seq with transcriptional profiling and bioinformatic approaches to define consensus DNA binding sequences proved to be effective for the elucidation of the regulon of the alternative σ factor SigX, revealing its role in complex virulence-associated phenotypes in P. aeruginosa.


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