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PMID: 8345519 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The minus 35-recognition region of Escherichia coli sigma 70 is inessential for initiation of transcription at an "extended minus 10" promoter.

Journal of molecular biology ·Vol. 232 ·No. 2 ·1993-07-20 ·Pages 406-18

Kumar A, Malloch RA, Fujita N, Smillie DA, Ishihama A, Hayward RS

Abstract

It is known that Escherichia coli promoters having the major minus 10 consensus sequence TATAAT, but lacking significant resemblance to consensus in the minus 35 region, allow transcriptional initiation in vivo and in vitro if they have an additional TGn motif immediately upstream of minus 10. To determine whether region 4.2 of sigma 70, whose normal role is sequence recognition at minus 35, is unnecessary for initiation of transcription at such "extended minus 10" promoters, we modified the sigma 70 gene so as to generate a carboxy-truncated polypeptide lacking the last 84 amino acids and therefore missing region 4.2. Our results show that both the intact and truncated sigma 70 allow purified RNA polymerase to initiate efficiently and specifically at an extended minus 10 promoter, whereas only the intact sigma 70 permits efficient initiation at normal promoters (defined by minus 35 and minus 10 hexamer sequences).

Related Genes
MeSH Terms
Bacterial Proteins/genetics,metabolism Base Sequence Chloramphenicol O-Acetyltransferase/biosynthesis DNA-Directed RNA Polymerases/genetics,metabolism Escherichia coli/genetics Genes, Bacterial/genetics Molecular Sequence Data Promoter Regions, Genetic/genetics Recombinant Fusion Proteins/biosynthesis Sequence Deletion Sequence Homology, Nucleic Acid Sigma Factor/genetics,metabolism Transcription, Genetic
Chemicals
Bacterial Proteins Recombinant Fusion Proteins Sigma Factor Chloramphenicol O-Acetyltransferase RNA polymerase sigma 70 DNA-Directed RNA Polymerases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kumar A
Institute of Cell and Molecular Biology, University of Edinburgh, Scotland, U.K.
Malloch R A
Fujita N
Smillie D A
Ishihama A
Hayward R S
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
0022-2836
Published
1993-07-20
Pages
406-18
Language
English
Region
England
NLM ID
2985088R
Subset
IM
Grants
Wellcome Trust · United Kingdom
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