Identification of DNA-binding proteins involved in regulation of expression of the Streptomyces aureofaciens sigF gene, which encodes sporulation sigma factor sigma(F).
Expression of the sigF gene encoding a sporulation-specific sigma factor, sigma(F), in Streptomyces aureofaciens is restricted only to sporulation. Gel mobility-shift assays using protein fractions from different developmental stages of S. aureofaciens revealed two different putative proteins specifically bound to the sigF promoter region: a protein (designated RsfA) present in young substrate mycelium, and a protein (designated RsfB) present in the course of sporulation. Based on the characteristic profiles of their appearance during differentiation, RsfA might be a repressor and RsfB an activator of sigF expression. The location of a specific binding site of the repressor-like protein (RsfA) was determined by gel mobility-shift assays of promoter deletion fragments and by DNase I footprinting analysis. The binding site mapped from nucleotides -87 to -25 relative to the transcription start point of the sigF promoter, and overlapped the -35 promoter region. Given the dependence of sigF expression upon whiH, the putative sporulation transcription factor WhiH was overproduced in Escherichia coli and used in the mobility-shift assays with the sigF promoter. However, no specific binding was detected, indicating an indirect dependence of sigF upon whiH.
Homerová
D
D
Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic1.
Ševčı Ková
B
B
Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic1.
Sprušanský
O
O
Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic1.
Kormanec
J
J
Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic1.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D012808:Sigma Factor / Q000235:genetics*; D013171:Spores, Bacterial / Q000235:genetics / Q000378:metabolism; D013304:Streptomyces aureofaciens / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C064971:FliA protein, Bacteria; D011994:Recombinant Proteins; D012097:Repressor Proteins; D012808:Sigma Factor; D014157:Transcription Factors; C403329:rsfA protein, Bacillus subtilis; C115233:whiH protein, Streptomyces; D003850:Deoxyribonuclease I