In vitro interactions of the histone-like protein IHF with the algD promoter, a critical site for control of mucoidy in Pseudomonas aeruginosa.
The ability of the histone-like element Integration Host Factor (IHF) to interact with the algD promoter was investigated. IHF from Escherichia coli was found to bind to the algD promoter and to form multiple protein-DNA complexes in gel mobility shift DNA binding assay. The highest affinity binding site for IHF was mapped by DNaseI footprinting analysis. This site spanned nucleotides -50 to -85 relative to the algD mRNA start site and overlapped a sequence matching the IHF consensus sequence WATCAANNNNTTR in 12 out of 13 base pairs. Previous studies have shown that deletion of sequences including a portion of this site adversely affects algD promoter activity. IHF binding to the algD promoter induced DNA bending. Western blot analysis with antibodies against E. coli IHF detected a cross-reactive protein of a similar molecular mass in Pseudomonas aeruginosa, suggesting the presence of an analogous factor in this organism.
Mohr
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Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
Deretic
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002237:Carbohydrate Dehydrogenases / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003851:Deoxyribonucleases; D005798:Genes, Bacterial; D006657:Histones / Q000378:metabolism*; D034982:Integration Host Factors; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D006657:Histones; D034982:Integration Host Factors; D012333:RNA, Messenger; C440503:integration host factor, Pseudomonas; D002237:Carbohydrate Dehydrogenases; C020562:GDPmannose dehydrogenase; D003851:Deoxyribonucleases