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Journal of Bacteriology, February 2000, p. 1181-1184, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Two Extracytoplasmic Function Sigma Subunits, sigma E and sigma FecI, of Escherichia coli: Promoter Selectivity and Intracellular Levels

Hiroto Maeda,dagger Miki Jishage, Tasuku Nomura, Nobuyuki Fujita, and Akira Ishihama*

National Institute of Genetics, Department of Molecular Genetics, Mishima, Shizuoka 411-5840, Japan

Received 30 August 1999/Accepted 17 November 1999

The promoter selectivity of two extracytoplasmic function (ECF) subfamily sigma  subunits, sigma E (sigma 24) and sigma FecI (sigma 18), of Escherichia coli RNA polymerase was analyzed by using an in vitro transcription system and various promoters. The Esigma E holoenzyme recognized only the known cognate promoters, rpoEP2, rpoHP3, and degP, and the Esigma FecI recognized only one known cognate promoter, fecA. The strict promoter recognition properties of sigma E and sigma FecI are similar to those of other minor sigma  subunits. Transcription by Esigma E and Esigma FecI was enhanced by high concentrations of glutamate, as in the case of other minor sigma  subunits. The optimum temperature for transcription by Esigma FecI was low, around 25°C, apparently in agreement with the high rate of iron sequestration by E. coli at low temperatures. By quantitative Western blot analysis, the intracellular levels of sigma E and sigma FecI in the uninduced steady-state culture of E. coli W3110 (type A) were determined to be 0.7 to 2.0 and 0.1 to 0.2 fmol per µg of total proteins (or 3 to 9 and 0.4 to 0.9 molecules per cell), respectively, and less than 1% of the level of the major sigma 70 subunit.

* Corresponding author. Mailing address: Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan. Phone: 81-559-81-6741. Fax: 81-559-81-6746. E-mail: aishiham{at}lab.nig.ac.jp.

dagger Permanent address: Kagoshima University, Faculty of Fisheries, Kagoshima 890-0056, Japan.

Journal of Bacteriology, February 2000, p. 1181-1184, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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