%0 Journal Article %A Homerová, D. %A Ševčı́ková, B. %A Sprušanský, O. %A Kormanec, J. %T Identification of DNA-binding proteins involved in regulation of expression of the Streptomyces aureofaciens sigF gene, which encodes sporulation sigma factor σF %D 2000 %J Microbiology, %V 146 %N 11 %P 2919-2928 %@ 1465-2080 %R https://doi.org/10.1099/00221287-146-11-2919 %K promoter %K repressor %K sigma factor %K differentiation %K Streptomyces aureofaciens %K tsp, transcription start point %I Microbiology Society, %X Expression of the sigF gene encoding a sporulation-specific sigma factor, σF, 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. %U https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-146-11-2919