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PMID: 1508713 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The promoter of the tgt/sec operon in Escherichia coli is preceded by an upstream activation sequence that contains a high affinity FIS binding site.

Nucleic acids research ·Vol. 20 ·No. 16 ·1992-08-25 ·Pages 4193-8

Slany RK, Kersten H

Abstract

The tgt/sec operon in E. coli consists of five genes: queA, tgt, ORF12, secD, and secF. QueA and Tgt participate in the biosynthesis of the hypermodified t-RNA nucleoside Queuosine, whereas SecD and SecF are involved in protein secretion. Examination of the promoter region of the operon showed structural similarity to promoter regions of the rrn-operons. An upstream activation sequence (UAS) containing a potential binding site for the factor of inversion stimulation (FIS) was found. Gel retardation assays and DNaseI footprinting indicated, that FIS binds specifically and with high affinity to a site centred at position -58. Binding of FIS caused bending of the DNA, as deduced from circular permutation analysis. Various 5' deletion mutants of the promoter region were constructed and fused to a lacZ reporter gene to determine the influence of the UAS element on the promoter strength. An approximately two-fold activation of the promoter by the UAS element was observed.

Related Genes
sec tgt
MeSH Terms
Base Sequence Binding Sites/genetics Carrier Proteins/genetics Consensus Sequence/genetics DNA Mutational Analysis DNA-Binding Proteins/genetics Escherichia coli/genetics Escherichia coli Proteins Factor For Inversion Stimulation Protein Integration Host Factors Molecular Sequence Data Nucleoside Q/genetics Operon/genetics Plasmids/genetics Promoter Regions, Genetic/genetics Recombinant Fusion Proteins/genetics
Chemicals
Carrier Proteins DNA-Binding Proteins Escherichia coli Proteins Factor For Inversion Stimulation Protein Integration Host Factors Recombinant Fusion Proteins integration host factor, E coli Nucleoside Q
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Slany R K
Institut für Biochemie, Universität Erlangen-Nürnberg, Germany.
Kersten H
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29 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/20.16.4193
Published
1992-08-25
Pages
4193-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334125
Subset
IM
Databases
GENBANK
M37702
External Links
PubMed source DOI: 10.1093/nar/20.16.4193 PMC Full Text View journal details
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