Differential binding of RNA polymerase to the pRM and pR promoters of bacteriophage lambda |
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Authors: | E M Owens G N Gussin |
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Affiliation: | Department of Zoology, University of Iowa, Iowa City, IA52242 U.S.A. Tel. 319-353-4663 |
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Abstract: | Escherichia coli RNA polymerase binding to the promoters pR and pRM of bacteriophage lambda was visualized and quantitated by electron microscopy. Although the two promoters are located close together in the phage genome, their proximity to the end of an 889-bp HaeIII DNA fragment made it possible to position binary complexes within 18 bp (2%) intervals. Thus, polymerase binding to pR and pRM could be distinguished by comparing the locations of binary complexes formed with wild-type and mutant (prm-) DNA at 37 degrees and 15 degrees C. We found that at 37 degrees C, RNA polymerase bound primarily to pR, while at 15 degrees C the efficiency of binding was the same at pRM as at pR. In addition, at 15 degrees C the overall efficiency of binding was significantly reduced relative to that at 37 degrees C. When the enzyme was incubated with prm- DNA, binding to pRM was reduced at both temperatures, as expected. Reduced binding to pRM was accompanied by an increase in binding to pR, apparently as a consequence of the low enzyme-to-DNA ratios used in these experiments. |
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Keywords: | Restriction fragments electron microscopy temperature affect closed and open binary complexes bp base pairs DTT dithiothreitol major rightward λ promoter SDS sodium dodecyl sulfate |
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