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PMID: 2990906 Published · ppublish English Journal Article

Regulation of the expression of the tufB operon: DNA sequences directly involved in the stringent control.

The EMBO journal ·Vol. 4 ·No. 4 ·1985-04-00 ·Pages 1053-8

Mizushima-Sugano J, Kaziro Y

Abstract

We have located the DNA sequence involved in the stringent control of the Escherichia coli tufB operon. Various deletion and insertion mutants of the promoter locus were constructed by in vitro mutagenesis, and their response to guanosine-5'-diphosphate-3'-diphosphate (ppGpp) was examined in a cell-free transcription system consisting of purified RNA polymerase holoenzyme. The nucleotide sequence (GpCpGpC) from positions -7 to -4 (designating the initiation site of mRNA as position +1) is responsible for the selective inhibition by ppGpp of tufB transcription. Point mutations were then constructed in which each one of the above four nucleotides was replaced by an A or T residue and tested for their response to ppGpp in the in vitro transcription system. The results indicated that the alteration of any nucleotide in the GpCpGpC sequence leads to the loss of the stringent response.

MeSH Terms
Base Sequence Chromosome Deletion DNA Restriction Enzymes Escherichia coli/enzymology,genetics GTP Pyrophosphokinase/genetics Genes Genes, Bacterial Guanosine Tetraphosphate/pharmacology Mutation Operon Phosphotransferases/genetics Transcription, Genetic/drug effects
Chemicals
Guanosine Tetraphosphate Phosphotransferases GTP Pyrophosphokinase DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Mizushima-Sugano J
Kaziro Y
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28 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1985-04-00
Pages
1053-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC554299
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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