Curation Information

Publication
Regulation of glutamate dehydrogenase expression in Pseudomonas putida results from its direct repression by NtrC under nitrogen-limiting conditions.;Hervás AB, Canosa I, Santero E;Molecular microbiology 2010 Oct; 78(2):305-19 [20735780]
TF
NtrC [Q88CY1, view regulon]
Reported TF sp.
Pseudomonas putida KT2440
Reported site sp.
Pseudomonas putida KT2440
Created by
Dinara Sagitova
Curation notes
-

Experimental Process

Promoter-lacZ fusion assays showed that NtrC controls gdhA expression in response to nitrogen availability. The transcription initiation site was identified by performing a primer extension analysis of the gdhA transcript. DNase I footprinting of the gdhA promoter region was used to identify the NtrC binding site. In vitro titration assays confirmed that NtrC directly represses the transcription of gdhA. Site-directed mutagenesis in conjunction with DNase I footprinting confirmed the binding specificity of the NtrC to its target site.

Transcription Factor Binding Sites


CGCCCTATTCCGGTGCG
CGCCCTATTCCGGTGCG

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
CGCCCTATTCCGGTGCG gdhA
... ... gdhA
Experimental technique details Beta-gal reporter assay - Experimental technique details DNAse footprinting (ECO:0005631) - Experimental technique details In-vitro transcription (ECO:0001204) - Experimental technique details Site directed mutagenesis (ECO:0005667) - repressor dimer