Free sigma factor of Escherichia coli RNA polymerase can bind to DNA

Abstract

Free sigma factor from E. coli RNA polymerase holoenzyme was shown to associate with supercoiled pBR350 and pBRH4 plasmid DNA s by two methods. The banding pattern of sigma factor through a nondenaturing polyacrylamide slab gel was significantly altered in the presence of supercoiled DNA; sigma factor had little or no affinity to linear, double- or single-stranded DNA. At saturation, approximately one sigma factor was bound/200 base pairs of supercoiled pBR350 DNA. By the nitrocellulose filter trapping method, sigma factor was able to bind supercoiled pBRH4 DNA much more efficiently than linear double-stranded pBRH4 DNA. These results suggest that sigma factor plays a role in the binding of holoenzyme to DNA and may be involved in locally denaturing DNA in the promoter region as postulated by previous investigators.