Curation Information

Publication
Requirements for Vibrio cholerae HapR binding and transcriptional repression at the hapR promoter are distinct from those at the aphA promoter.;Lin W, Kovacikova G, Skorupski K;Journal of bacteriology 2005 May; 187(9):3013-9 [15838027]
TF
HapR [A0A0H3Q915, view regulon]
Reported TF sp.
Vibrio cholerae C6706
Reported site sp.
Vibrio cholerae C6706
Created by
Erill Lab
Curation notes
-

Experimental Process

Lux reporter assays showed repression of chromosomal hapR by plasmid-borne HapR. RACE PCR was used to determine the transcription start site. Binding of purified HapR to its promoter was demonstrated with EMSA, using multiple fragment sizes, and further corroborated with DNAse footprinting to define the site, identified by similarity to known motif. Site directed mutagenesis was used to identify mutations that prevent binding.

Transcription Factor Binding Sites


CAAATAACAAAATAATCATTAG
CAAATAACAAAATAATCATTAG

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
CAAATAACAAAATAATCATTAG VCD_001025,
... ... VCD_001025 VCD_001024
Experimental technique details Consensus search (ECO:0005624) - Experimental technique details DNAse footprinting (ECO:0005631) - Experimental technique details EMSA (ECO:0001807) - Experimental technique details Luciferase reporter assay (ECO:0005648) - Experimental technique details RACE PCR (ECO:0005661) - Experimental technique details Site directed mutagenesis (ECO:0005667) - repressor not specified