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J. Biochem, 1995, Vol. 118, No. 3 488-493
© 1995 Japanese Biochemical Society

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Purification and Characterization of RNA Polymerase Holoenzyme (EóB) from Vegetative-Phase Mycelia of Streptomyces griseus

Hidenori Shinkawa*,1, Nobuyuki Fujita{dagger}, Tetsuo Shiina{ddagger}, Kan Tanaka{ddagger}, Hideo Takahashi{ddagger}, Akira Ishihama{dagger} and Osamu Nimi*

*Department of Fermentation Technology, Faculty of Engineering, Hiroshima University Higashi-Hiroshima, Hiroshima 739
{dagger}Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411
{ddagger}Institute of Molecular and Cellular Bioscience, The University of Tokyo Bunkyo-ku, Tokyo 110

1 To whom correspondence should be addressed. Phone: + 81-824-24-7768, Fax:+81-824-22-7191, e-mail: hshinka{at}ue.ipc.hiroshima-u.ac.jp

RNA polymerase was purified from vegetative-phase mycelia of Streptomyces griseus by a series of ion-exchange chromatographies. By western blot analysis using antiserum against S. coelicolor HrdB, which is a principal sigma factor (óhrdB), the purified holoenzyme was found to contain óB (= óhrdB) of S. griseus. Significant amounts of HrdB protein were, however, eluted from the DEAE column at lower concentrations of KCl than that required for elution of the holoenzyme containing óH, suggesting that óB is dissociated from the core enzyme, or an excess amount of óB exists in S. griseus cells. The holoenzyme containing óB (Eón) transcribed in vitro the dagA promoter of S. coelicolor, and the hrdB and hsp70 promoters of 5. griseus, suggesting that it is involved in transcription of the essential genes. EóB may be a major form of RNA polymerase holoenzyme in the growing phase of S. griseus.


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