Nucleic Acids Research, Vol 25, Issue 13 2566-2573, Copyright © 1997 by Oxford University Press
JG Kang, MY Hahn, A Ishihama and JH Roe
We examined the promoter selectivity of RNA polymerase (RNAP) from
Streptomyces coelicolor at two growth phases by in vitro transcription.
Distinct sets of promoters were preferentially recognized by either
exponential or stationary phase RNAP. No change in molecular weight or net
charge of the core subunits was observed, suggesting that the associated
specificity factors determined phase-specific promoter selectivity of the
holoenzyme. Five different specificity factors and their cognate promoters
were identified by in vitro holoenzyme reconstitution and transcription
assays. sigma66 (sigma hrdB) and sigma46 (sigma hrdD) recognized promoters
(rrnD p2 and dagA p4 for sigma66, actII-orf4 p and whiB p2 for sigma46)
preferentially transcribed by the exponential phase RNAP. sigma52
recognized promoters (dagA p3 and actIII px1) preferentially transcribed by
the stationary phase RNAP. Sigma28 (sigma sigE) recognized promoters (hrdD
p1, whiB p1 and dagA p2) transcribed equally by both RNAPs. A novel 31 kDa
specificity factor recognized actIII px2, glnR p2 and hrdD p2 promoters
preferentially transcribed by the stationary phase RNAP. This factor was
isolated from the stationary phase RNAP and reconstituted holoenzyme in
vitro as a sigma factor. The N-terminal sequence suggests that it is a
novel factor. By examining phase-specific promoter recognition pattern we
can predict that holoenzyme Esigma52 and Esigma31 activities are higher in
the stationary phase, whereas Esigma66 and Esigma46activities are higher in
the exponential phase. Possible promoter sequences recognized by some of
these sigma factors were suggested.
ARTICLES
Identification of sigma factors for growth phase-related promoter selectivity of RNA polymerases from Streptomyces coelicolor A3(2)
Department of Microbiology, College of Natural Sciences, and Research Center for Molecular Microbiology, Seoul National University, Seoul 151- 742, Korea.
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