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Mode of promoter recognition by the Escherichia coli RNA polymerase holoenzyme containing the σ s subunit: identification of the recognition sequence of the fic promoter
Author(s) -
Hiratsu Keiichiro,
Shinagawa Hideo,
Makino Kozo
Publication year - 1995
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1995.18050841.x
Subject(s) - biology , rna polymerase , recognition sequence , escherichia coli , protein subunit , genetics , identification (biology) , specificity factor , promoter , microbiology and biotechnology , polymerase , sequence (biology) , gene , computational biology , gene expression , restriction enzyme , botany
Transcription of a number of genes during stationary phase in Escherichia coli requires the alternative sigma factor σ s , encoded by the rpoS gene. No consensus sequence for the promoters recognized by the RNA polymerase holoenzyme containing the σ s subunit (Eσ s ) is known. To identify the Eσ s recognition sequence, we analysed the promoter region of the fic gene, whose expression depends on Eσ s both in vivo and in vitro . By random mutagenesis, we isolated a large number of the fic promoter mutants that had defective promoter activities in an rpoS,+ strain. All such mutants contained alterations within the −10 region, TATACT, whereas no mutant was isolated with alterations in the −35 region. Based on further analyses including scanning mutagenesis and DNase 1 footprinting assays of the fic promoter region, and in vitro transcription assays, we conclude that the −10 hexamer is essential to transcription initiation from the fie promoter by Eσ s .