Expression, isolation and properties of Fur (ferric uptake regulation) protein ofEscherichia coli K 12

Wee, Sechan; Neilands, J. B.; Bittner, Michael; Hemming, Bruce C.; Haymore, Barry L.; Seetharam, Ramnath

doi:10.1007/bf01128019

Cited by 77 publications

(92 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The probe's migration through a polyacrylamide gel was retarded after adding increasing concentrations of Fur (figure 4A), indicating that Fur bound the iha promoter region. Multiple shift products were observed, as has been described for Fur [25,26]. Abundance of the higher molecular weight complex was decreased when the binding reaction was incubated in increasing (competitive) concentrations of unlabeled probe or of a short double-stranded oligonucleotide consisting of only the putative iha Fur-binding site flanked by 7 bp on either side ( figure 4C).…”

Section: Interaction Of the Iha Promoter With Furmentioning

confidence: 86%

See 1 more Smart Citation

Expression of the Escherichia coli IrgA homolog adhesin is regulated by the ferric uptake regulation protein

Rashid

Tarr

Moseley

2006

Microbial Pathogenesis

View full text Add to dashboard Cite

The IrgA homologue adhesin (Iha) is an adherence-conferring outer membrane protein of Escherichia coli associated with enterohemorrhagic and uropathogenic strains. Here, we used primer extension analysis to identify iha promoters in O157:H7 and uropathogenic E. coli strains. Transcriptional fusions demonstrated that iha transcription is repressed by iron. Gel shifts using purified ferric uptake regulator protein (Fur) demonstrated that repression involves a direct interaction between Fur and the iha promoter. We identified strain-dependent differences in iha expression and determined that single nucleotide polymorphisms upstream of the iha promoter, in particular position −85, contribute to differences in expression levels.

show abstract

Section: Interaction Of the Iha Promoter With Furmentioning

confidence: 86%

“…pET21dFur was transformed into BL21(DE3) (Novagen), and the purification strategy was based on previously published methods [48,25,26]. An overnight culture was diluted 1:100 and grown to OD 600 = 0.6 while shaking at 37°C.…”

Section: Fur Purificationmentioning

confidence: 99%

Expression of the Escherichia coli IrgA homolog adhesin is regulated by the ferric uptake regulation protein

Rashid

Tarr

Moseley

2006

Microbial Pathogenesis

View full text Add to dashboard Cite

show abstract

“…This value is relatively high as compared with that of E. coli Fur to the iucA promoter (5 nM; Ref. 28) and the sodA promoter (10 -20 nM; Ref. 29).…”

Section: Determination Of the Transcription Start Site For Fatdcba Mrmentioning

confidence: 96%

“…Based on in vitro data it has been suggested that the E. coli Fur protein is active as a dimer or as a multimer (17,18,28) that binds to its 19-bp dyad symmetrical operator sequence, so called Fur consensus sequence or iron box (19, 28; Fig. 7) in the promoter region.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the Interaction between Fur and the Iron Transport Promoter of the Virulence Plasmid in Vibrio anguillarum

Chai

Welch

Crosa

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

The expression of iron transport genes fatDCBA in Vibrio anguillarum strain 775 is negatively regulated by two iron-responsive repressors, the Fur protein and the antisense RNA, RNA␣. Here we report the identification of the promoter for the iron transport genes and studied the interaction between the V. anguillarum Fur protein and this promoter. The iron transport promoter was localized in a region approximately 300 base pairs upstream of fatD by both primer extension and S1 mapping analysis. High activity of the promoter was measured in response to iron depletion in the wild-type strain when a promoter-lacZ fusion was examined, whereas the promoter was constitutive in the Fur-deficient strain. Gel retardation and DNase I footprint analysis showed that Fur binds specifically to two contiguous sites comprising the promoter region and the region downstream of the transcription start site. The identified Fur binding sites showed a low degree of homology to each other as well as to the consensus sequence for the Escherichia coli Fur protein. DNase I footprints pattern suggested a sequential interaction of Fur with these two sites that renders a protection in the template strand and a hypersensitivity to the nuclease in the nontemplate strand.

show abstract

“…In addition to this, Fur seems to polymerize around the DNA of the aerobactin promoter with a precise directionality but unclear stoichiometry. Furthermore, one study by Wee et al (1988) suggested that Fur could form tripartite complexes with the target DNA sequences and the RNA polymerase (RNAP).…”

Section: Introductionmentioning

confidence: 99%

Metalloregulation in vitro of the aerobactin promoter of Escherichia coli by the Fur (ferric uptake regulation) protein

Escolar

Lorenzo

Pérez-Martı́n

1997

Molecular Microbiology

View full text Add to dashboard Cite

SummaryThe mechanism of transcriptional repression of the aerobactin operon of Escherichia coli by the Fe 2þ -responsive Fur (ferric uptake regulation) protein has been investigated. In the presence of a divalent metal, such as Mn 2þ , the Fur protein sequentially occupies two defined sites at the aerobactin promoter region, followed by a looser occupation of upstream DNA sequences. However, binding to the primary target site suffices for the entire repression effect. Comparison of transcription patterns generated with run-off experiments in the presence and absence of heparin showed that access of the RNA polymerase to the principal ¹35/¹10 hexamers of the promoter region was fully prevented by Fur-Mn 2þ bound to its primary site. Similarly, promoter-bound RNA polymerase could not be competed out from the DNA even in the presence of a large Fur-Mn 2þ excess, although the repressor could immediately bind its target sequence at the region as soon as RNA polymerase moved away from the promoter during transcription. The high affinities of either protein for the promoter produce, in practice, a first-come, first-served effect that helps the system to respond instantly to changes in the iron status of the cells.

show abstract

Expression, isolation and properties of Fur (ferric uptake regulation) protein ofEscherichia coli K 12

Cited by 77 publications

References 22 publications

Expression of the Escherichia coli IrgA homolog adhesin is regulated by the ferric uptake regulation protein

Expression of the Escherichia coli IrgA homolog adhesin is regulated by the ferric uptake regulation protein

Characterization of the Interaction between Fur and the Iron Transport Promoter of the Virulence Plasmid in Vibrio anguillarum

Metalloregulation in vitro of the aerobactin promoter of Escherichia coli by the Fur (ferric uptake regulation) protein

Contact Info

Product

Resources

About