Nucleic Acids Research, 2000, Vol. 28, No. 18 3497-3503
© 2000 Oxford University Press
Competition among seven Escherichia coli 
subunits: relative binding affinities to the core RNA polymerase
1National Institute of Genetics, Department of Molecular Genetics, Mishima, Shizuoka 411-8540, Japan and 2Kagoshima University, Faculty of Fisheries, Kagoshima 890-0056, Japan
Seven different species of the RNA polymerase 
subunit exist in Escherichia coli, each binding to a single species of the core enzyme and thereby directing transcription of a specific set of genes. To test the competition model in the global regulation of gene transcription, all seven E.coli subunits have been purified and compared for their binding affinities to the same core RNA polymerase (E). In the presence of a fixed amount of 70, the principal for growth-related genes, the level of E70 holoenzyme formation increased linearly with the increase in core enzyme level, giving an apparent Kd for the core enzyme of 0.26 nM. Mixed reconstitution experiments in the presence of a fixed amount of core enzyme and increasing amounts of an equimolar mixture of all seven subunits indicated that 70 is strongest in terms of core enzyme binding, followed by N, F, E/FecI, H and S in decreasing order. The orders of core binding affinity between 70 and N and between 70 and H were confirmed by measuring the replacement of one core-associated by another subunit. Taken together with the intracellular levels, we tried to estimate the number of each holoenzyme form in growing E.coli cells.
* To whom correspondence should be addressed. Tel: +81 559 81 6741; Fax: +81 559 81 6746; Email: aishiham@lab.nig.ac.jp
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