| pubmed-article:7306497 | pubmed:abstractText | RNA polymerase (RNA nucleotidyltransferase, EC 2.7.7.6) of Agrobacterium tumefaciens has been purified according to a fast and efficient procedure. The method involves only two chromatographic steps and yields a highly active enzyme. The RNA polymerase was studied with respect to the ability to bind its homologous genome. A. tumefaciens deoxyribonucleic acid (DNA) binds the enzyme even when fragmented at undergenic size (300 base pairs). The general binding is unspecific and very labile at low concentrations of heparin (0.66 micrograms/mL). The number and distribution of the stable binding sites, class A sites [Hinkle, D., & Chamberlin, M. J. (1972) J. Mol. Biol. 70, 157-185], have been calculated from the heparin-induced dissociation kinetics of binary complexes formed between the enzyme and DNA fragments of various sizes. A total of 3.5 x 10(3) class A sites (forming binary complexes with a half-life of 16.6 min) are present on A. tumefaciens genome, a large number of which show a distribution of 800-1000 base pairs. The rest have a more widely spaced distribution. The interactions between Escherichia coli RNA polymerase and the A. tumefaciens template have also been examined, and it has been observed that E. coli holoenzyme forms stable complexes with a shorter half-life and recognizes a lower number of class A sites on A. tumefaciens genome. | lld:pubmed |
