The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.

The promoter selectivity of Escherichia coli RNA polymerase is determined by the sigma subunit with promoter recognition activity. The model prokaryote Escherichia coli contains seven species of the sigma subunit, each recognizing a specific set of promoters. The major sigma subunit, sigma-70 encode...

Full description

Bibliographic Details
Main Authors: Tomohiro Shimada, Yukiko Yamazaki, Kan Tanaka, Akira Ishihama
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3946193?pdf=render
_version_ 1819092631515299840
author Tomohiro Shimada
Yukiko Yamazaki
Kan Tanaka
Akira Ishihama
author_facet Tomohiro Shimada
Yukiko Yamazaki
Kan Tanaka
Akira Ishihama
author_sort Tomohiro Shimada
collection DOAJ
description The promoter selectivity of Escherichia coli RNA polymerase is determined by the sigma subunit with promoter recognition activity. The model prokaryote Escherichia coli contains seven species of the sigma subunit, each recognizing a specific set of promoters. The major sigma subunit, sigma-70 encoded by rpoD, plays a major role in transcription of growth-related genes. Concomitant with the increase in detection of promoters functioning in vivo under various stressful conditions, the variation is expanding in the consensus sequence of RpoD promoters. In order to identify the canonical sequence of "constitutive promoters" that are recognized by the RNA polymerase holoenzyme containing RpoD sigma in the absence of supporting transcription factors, an in vitro mixed transcription assay was carried out using a whole set of variant promoters, each harboring one base replacement, within the model promoter with the conserved -35 and -10 sequences of RpoD promoters. The consensus sequences, TTGACA(-35) and TATAAT(-10), were identified to be ideal for the maximum level of open complex formation and the highest rate of promoter opening, respectively. For identification of the full range of constitutive promoters on the E. coli genome, a total of 2,701 RpoD holoenzyme-binding sites were identified by Genomic SELEX screening, and using the reconfirmed consensus promoter sequence, a total of maximum 669 constitutive promoters were identified, implying that the majority of hitherto identified promoters represents the TF-dependent "inducible promoters". One unique feature of the constitutive promoters is the high level of promoter sequence conservation, about 85% carrying five-out-of-six agreements with -35 or -10 consensus sequence. The list of constitutive promoters provides the community resource toward estimation of the inducible promoters that operate under various stressful conditions in nature.
first_indexed 2024-12-21T22:58:41Z
format Article
id doaj.art-0158dbf8b46a48048de9efbd98abc1ba
institution Directory Open Access Journal
issn 1932-6203
language English
last_indexed 2024-12-21T22:58:41Z
publishDate 2014-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj.art-0158dbf8b46a48048de9efbd98abc1ba2022-12-21T18:47:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0193e9044710.1371/journal.pone.0090447The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.Tomohiro ShimadaYukiko YamazakiKan TanakaAkira IshihamaThe promoter selectivity of Escherichia coli RNA polymerase is determined by the sigma subunit with promoter recognition activity. The model prokaryote Escherichia coli contains seven species of the sigma subunit, each recognizing a specific set of promoters. The major sigma subunit, sigma-70 encoded by rpoD, plays a major role in transcription of growth-related genes. Concomitant with the increase in detection of promoters functioning in vivo under various stressful conditions, the variation is expanding in the consensus sequence of RpoD promoters. In order to identify the canonical sequence of "constitutive promoters" that are recognized by the RNA polymerase holoenzyme containing RpoD sigma in the absence of supporting transcription factors, an in vitro mixed transcription assay was carried out using a whole set of variant promoters, each harboring one base replacement, within the model promoter with the conserved -35 and -10 sequences of RpoD promoters. The consensus sequences, TTGACA(-35) and TATAAT(-10), were identified to be ideal for the maximum level of open complex formation and the highest rate of promoter opening, respectively. For identification of the full range of constitutive promoters on the E. coli genome, a total of 2,701 RpoD holoenzyme-binding sites were identified by Genomic SELEX screening, and using the reconfirmed consensus promoter sequence, a total of maximum 669 constitutive promoters were identified, implying that the majority of hitherto identified promoters represents the TF-dependent "inducible promoters". One unique feature of the constitutive promoters is the high level of promoter sequence conservation, about 85% carrying five-out-of-six agreements with -35 or -10 consensus sequence. The list of constitutive promoters provides the community resource toward estimation of the inducible promoters that operate under various stressful conditions in nature.http://europepmc.org/articles/PMC3946193?pdf=render
spellingShingle Tomohiro Shimada
Yukiko Yamazaki
Kan Tanaka
Akira Ishihama
The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
PLoS ONE
title The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
title_full The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
title_fullStr The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
title_full_unstemmed The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
title_short The whole set of constitutive promoters recognized by RNA polymerase RpoD holoenzyme of Escherichia coli.
title_sort whole set of constitutive promoters recognized by rna polymerase rpod holoenzyme of escherichia coli
url http://europepmc.org/articles/PMC3946193?pdf=render
work_keys_str_mv AT tomohiroshimada thewholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT yukikoyamazaki thewholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT kantanaka thewholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT akiraishihama thewholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT tomohiroshimada wholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT yukikoyamazaki wholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT kantanaka wholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli
AT akiraishihama wholesetofconstitutivepromotersrecognizedbyrnapolymeraserpodholoenzymeofescherichiacoli