Mutational analysis of upstream sequences required for transcriptional activation of the Klebsiella pneumoniae nifH promoter.

Nucleic acids research ·Vol. 15 ·No. 23 ·1987-12-10 ·Pages 9945-56

Buck M, Cannon W, Woodcock J

Abstract

Upstream sequences of the Klebsiella pneumoniae nifH promoter were mutagenised and activation of the mutated promoters by the nif-specific transcriptional activator protein NifA examined in vivo. Of the sixteen mutations analysed, only those within the nifH upstream activator sequence (UAS), characterised by a TGT-N10-ACA motif, influenced nifH promoter activity. Mutations altering the two-fold rotational symmetry of the UAS or the spacing between the TGT and ACA motifs reduced promoter activity, consistent with the UAS functioning as a NifA binding site. The bases flanking the TGT-ACA motif of the UAS also appear to influence activation by NifA. Substituting the nifH UAS with a binding site for the transcriptional activator NtrC resulted in improved NtrC-dependent activation of the nifH promoter demonstrating that the activator specificity of the nifH promoter is dependent upon the presence of the appropriate upstream sequences to which the activator binds.

MeSH Terms

Base Sequence DNA Mutational Analysis/methods Genes, Bacterial Klebsiella pneumoniae/genetics Nitrogen Fixation/genetics Nucleic Acid Conformation Phenotype Promoter Regions, Genetic Transcription, Genetic

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Buck M

AFRC IPSR, University of Sussex, Brighton, UK.

Cannon W

Woodcock J

References (21)

21 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/15.23.9945

Published

1987-12-10

Pages

9945-56

Language

English

Region

England

NLM ID

0411011

PMCID

PMC306542

Subset

External Links

PubMed source DOI: 10.1093/nar/15.23.9945 PMC Full Text View journal details

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Mutational analysis of upstream sequences required for transcriptional activation of the Klebsiella pneumoniae nifH promoter.

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