Promoter selectivity of the Bacillus subtilis RNA polymerase sigmaA and sigmaH holoenzymes

Journal of Biochemistry
M Fujita, Y Sadaie

Abstract

The sigmaH of Bacillus subtilis directs transcription of a large number of early sporulation genes, whereas the principal sigma factor, sigmaA, is essential for the transcription of the genes for vegetative growth and early sporulation. We have purified sigmaA and sigmaH proteins, and characterized their properties. The genes encoding sigmaA or sigmaH were separately cloned into an expression vector under the control of T7 promoter. Both proteins were overproduced in Escherichia coli BL21(DE3) and purified from inclusion bodies after solubilization with guanidine hydrochloride. Antigenicities and N-terminal amino acid sequences of the overproduced proteins were used to identify both proteins. Unlike sigmaA protein, sigmaH protein showed a DNA-binding ability. To compare the promoter selectivity of the sigmaA protein with that of the sigmaH protein, transcription in vitro of 16 promoters was performed using RNA polymerase holoenzymes reconstituted from a purified core enzyme with either sigmaH or sigmaA. These holoenzymes correctly recognized each of the cognate promoters; sigmaH-RNA polymerase recognized sigmaH promoters but not sigmaA promoters, and vice versa. A competition experiment for core RNA polymerase using sigmaA and ...Continue Reading

Citations

Sep 20, 2011·Journal of Bacteriology·Prahathees EswaramoorthyMasaya Fujita
Jul 3, 2002·Bioscience, Biotechnology, and Biochemistry·Daisuke FukudaAyaaki Ishizaki
Aug 6, 2011·The Journal of General and Applied Microbiology·Koichi YanoKei Asai
May 25, 2010·FEMS Microbiology Reviews·Umender K Sharma, Dipankar Chatterji
Sep 8, 2009·FEMS Microbiology Letters·Rita A EckartAndreas Licht
Feb 15, 2000·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·M Fujita

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