Curation Information

Publication
Identification and characterization of the functional toxboxes in the Vibrio cholerae cholera toxin promoter.;Dittmer JB, Withey JH;Journal of bacteriology 2012 Oct; 194(19):5255-63 [22821976]
TF
ToxT [A5F384, view regulon]
Reported TF sp.
Vibrio cholerae O395
Reported site sp.
Vibrio cholerae O395
Created by
Elliot White
Curation notes
-

Experimental Process

Citing previous studies as evidence of ToxT control of the ctxAB operon, researchers first ran a CP footprinting assay, which showed protection of the latter 3 members of the GATTTTT heptad repeat that is canonically associated with ToxT and ToxR binding. Researchers then conducted B-gal assays of point mutations of the positions of the footprint. Finally, researchers ran EMSAs different length promoter fragments. All three experiments suggested the presence of 2 ToxT binding sites covering repeats 5 and 6, and a portion of the AT-rich region after the repeats.

Transcription Factor Binding Sites


GATTTTTGATTTT
GATTTCAAATAAT
GATTTTTGATTTT
GATTTCAAATAAT

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
GATTTTTGATTTT ctxA,
... ... ctxA zot ace VC0395_0509 cep rstB2 rstA1 VC0395_0505
Experimental technique details Beta-gal reporter assay - Experimental technique details Copper-phenanthroline footprinting (ECO:0005626) - Experimental technique details EMSA (ECO:0001807) - Experimental technique details Site directed mutagenesis (ECO:0005667) - repressor monomer
GATTTCAAATAAT ctxA,
... ... ctxA zot ace VC0395_0509 cep rstB2 rstA1 VC0395_0505
Experimental technique details Beta-gal reporter assay - Experimental technique details Copper-phenanthroline footprinting (ECO:0005626) - Experimental technique details EMSA (ECO:0001807) - Experimental technique details Site directed mutagenesis (ECO:0005667) - repressor monomer