Curation Information

Publication: The svpA-srtB locus of Listeria monocytogenes: fur-mediated iron regulation and effect on virulence.;Newton SM, Klebba PE, Raynaud C, Shao Y, Jiang X, Dubail I, Archer C, Frehel C, Charbit A;Molecular microbiology 2005 Feb; 55(3):927-40 [15661014]
TF: Fur [Q8Y5U9, view regulon]
Reported TF sp.: Listeria monocytogenes EGD-e
Reported site sp.: Listeria monocytogenes EGD-e
Created by: Erill Lab
Curation notes: -

Experimental Process

The authors identify a putative Fur-box in the promoter region of lmo2186, and confirm with RT-PCR that this gene forms a single transcriptional unit with seven other genes (the svpA-srtB cluster, lmo2186-lmo2180). They verify using a Fur mutant and iron supplementation that Fur acts as a repressor of this operon via RT-PCR, Western immunoblotting and a GFP fusion. They further characterized the influence of this regulation in virulence phenotypes for L. monocytogenes.

Transcription Factor Binding Sites

Sites reported in the paper
Sites matched in the genome

GACAATGATAATCATTATC

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
GACAATGATAATCATTATC	lmo2180, lmo2181, lmo2182, lmo2183, lmo2184, lmo2185, lmo2186		Experimental technique details Ad-hoc quantitative phenotype observation (ECO:0005676) Ad-hoc quantitative phenotype observation - ECO:0005676 A quantifiable phenotypic trait (e.g. glucose intake) is assessed in a quantitative manner after induction of the regulatory mechanism and used as a natural reporter.. The observable phenotypic trait should be described in the experimental notes. - Experimental technique details GFP Promoter Fusion (ECO:0005636) GFP Promoter Fusion - ECO:0005636 Target promoter is inserted in front of a GFP gene, and the target TF is induced. Expression levels of GFP should thereby depend on the target TF. - Experimental technique details qRT-PCR [RNA] (ECO:0001808) qRT-PCR [RNA] - ECO:0001808 Quantitative Reverse-Transcription PCR is a modification of PCR in which RNA is first reverse transcribed into cDNA and this is amplified measuring the product (qPCR) in real time. It therefore allows one to analyze transcription by directly measuring the product (RNA) of a gene's transcription. If the gene is transcribed more, the starting product for PCR is larger and the corresponding volume of amplification is also larger. - Experimental technique details Visual sequence inspection (nan) Visual sequence inspection - nan Visual inspection of promoter region to identify putative binding sites based on previous knowledge - Experimental technique details Western blot (quantitative) expression analysis (ECO:0000279) Western blot (quantitative) expression analysis - ECO:0000279 A Western blot (also frequently called immunoblot) is a gel-based technique used to determine the presence of specific proteins in a sample by means of separation (through gel electrophoresis) and detection (using a protein-specific antibody). Less frequent than the equivalent RNA Northern blot, it is typically used in TF-binding site analysis to validate differential regulation of target genes. -	repressor	not specified