分子生物学Chapter 9 DNA–Protein课件.pptVIP

Effect of DNA Looping on DNase Susceptibility Operators separated by Integral number of double-helical turns can loop out DNA to allow cooperative binding Nonintegral number of turns requires proteins to bind to opposite faces of DNA and no cooperative binding When λ operators are separated by an integral number of helical turns, the DNA in between can loop out to allow cooperative binding. When the operators are separated by a nonintegral number of helical turns, the proteins have to bind to opposite faces of the DNA double helix, so no cooperative binding can take place. SUMMARY 3) Enhancers σ70 σ54(σN) (nitrogen starvation) Enhancers are nonpromoter DNA elements that bind protein factors and stimulate transcription Can act at a distance Originally found in eukaryotes Recently found in prokaryotes NtrC--enhancer 70 bp How does the enhancer interact with the promoter? The enhancer can still function even if it and the promoter are on separate DNA molecules, as long as the two molecules are linked in a catenane 2) The enhancer has to be at least 70 bp away from the promoter to perform its function. Plot of the loop lengths. The investigators measured the lengths of several loops and plotted their lengths versus the number of loops of a given length. The peak occurred at 390 bp, which is exactly the spacing between the enhancer and the promoter. SUMMARY The E. coli glnA gene is an example of a prokaryotic gene that depends on an enhancer for its transcription. The enhancer binds the NtrC protein, which interacts with polymerase bound to the promoter at least 70 bp away. Hydrolysis of ATP by NtrC allows the formation of an open promoter complex so transcription can take place. The two proteins appear to interact by looping out the DNA in between. The phage T4 late enhancer is mobile; it is part of the phage DNA-replication apparatus. Because this enhancer must be on the same DNA molecule as the late promoters, it probably does not act by DNA looping. Prok