Curation Information

Publication
Multiple-level regulation of 2,4-diacetylphloroglucinol production by the sigma regulator PsrA in Pseudomonas fluorescens 2P24.;Wu X, Liu J, Zhang W, Zhang L;PloS one 2012; 7(11):e50149 [23209661]
TF
PsrA [Q4KFB4, view regulon]
Reported TF sp.
Pseudomonas fluorescens 2P24
Reported site sp.
Pseudomonas fluorescens 2P24
Created by
Dinara Sagitova
Curation notes
-

Experimental Process

A sequence alignment of the phlA gene from five 2,4-DAPG producing Pseudomonas strains revealed a very well conserved palindromic sequence GAAACN5GTTTC. EMSA verified binding of PsrA to the phlA promoter. Site-directed mutagenesis confirmed that binding was abolished when mutations were introduced into the putative PsrA binding site. phlA-lacZ transcriptional fusion in wild-type and psrA mutant strains confirmed that PsrA represses phlA expression. Western blot assay showed that levels of PhlA protein increased in the psrA mutant compared to the wild type strain.

Transcription Factor Binding Sites


GAAACGTATGTTTC
CAAACAAGTGTTTG
GAAACGTATGTTTC
CAAACAAGTGTTTG

Gene Regulation

Regulated genes for each binding site are displayed below. Gene regulation diagrams show binding sites, positively-regulated genes, negatively-regulated genes, both positively and negatively regulated genes, genes with unspecified type of regulation. For each indvidual site, experimental techniques used to determine the site are also given.

Site sequence Regulated genes Gene diagram Experimental techniques TF function TF type
GAAACGTATGTTTC psrA,
... ... psrA lexA sulA
Experimental technique details Beta-gal reporter assay - Experimental technique details EMSA (ECO:0001807) - Experimental technique details Multiple sequence alignment (MSA) (ECO:0005556) - Experimental technique details Site directed mutagenesis (ECO:0005667) - Experimental technique details Western blot (quantitative) expression analysis (ECO:0000279) - repressor not specified
CAAACAAGTGTTTG psrA,
... ... psrA lexA sulA
Experimental technique details Beta-gal reporter assay - Experimental technique details EMSA (ECO:0001807) - Experimental technique details Multiple sequence alignment (MSA) (ECO:0005556) - Experimental technique details Site directed mutagenesis (ECO:0005667) - Experimental technique details Western blot (quantitative) expression analysis (ECO:0000279) - repressor not specified