Integration host factor (IHF) modulates the expression of the pyrimidine-specific promoter of the carAB operons of Escherichia coli K12 and Salmonella typhimurium LT2

Molecular & General Genetics : MGG
Daniel CharlierN Glansdorff

Abstract

We report the identification of Integration Host Factor (IHF) as a new element involved in modulation of P1, the upstream pyrimidine-specific promoter of the Escherichia coli K12 and Salmonella typhimurium carAB operons. Band-shift assays, performed with S-30 extracts of the wild type and a himA, hip double mutant or with purified IHF demonstrate that, in vitro, this factor binds to a region 300 bp upstream of the transcription initiation site of P1 in both organisms. This was confirmed by deletion analysis of the target site. DNase I, hydroxyl radical and dimethylsulphate footprinting experiments allowed us to allocate the IHF binding site to a 38 bp, highly A+T-rich stretch, centred around nucleotide -305 upstream of the transcription initiation site. Protein-DNA contacts are apparently spread over a large number of bases and are mainly located in the minor groove of the helix. Measurements of carbamoyl-phosphate synthetase (CPSase) and beta-galactosidase specific activities from car-lacZ fusion constructs of wild type or IHF target site mutants introduced into several genetic backgrounds affected in the himA gene or in the pyrimidine-mediated control of P1 (carP6 or pyrH+/-), or in both, indicate that, in vivo, IHF influence...Continue Reading

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Citations

Nov 20, 2002·Research in Microbiology·Pedro Miguel SantosElisabetta Zennaro
Sep 13, 2000·Nucleic Acids Research·L FloreaM McClelland
May 18, 2010·Nucleic Acids Research·Phu Le Minh NguyenDaniel Charlier
Sep 6, 2000·Journal of Molecular Biology·D CharlierN Glansdorff
Nov 10, 2017·International Journal of Genomics·Jesús Espinal-EnríquezEnrique Hernández-Lemus
Dec 1, 2004·EcoSal Plus·Daniel Charlier, Nicolas Glansdorff
Jun 1, 1995·Microbiological Reviews·K E SandersonK E Rudd
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·M K Berlyn

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