Prediction of the pho regulon in Streptomyces clavuligerus DSM 738

Negative control of clavulanic acid by phosphate in Streptomyces clavuligerus DSM 738 suggests that a pho regulon may exist in this bacterium. S. clavuligerus PhoP was expressed with a C-terminal His-tag in Escherichia coli and purified. Binding of PhoP-His 6 to promoter fragments of phoRP/phoU and...

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Bibliographic Details
Published in:The New microbiologica 2012-10, Vol.35 (4), p.447-457
Main Authors: Salehghamari, Ensieh, Hamedi, Javad, Elahi, Elahe, Sepehrizadeh, Zargham, Sadeghi, Mehdi, Muth, Guenther
Format: Article
Language:English
Subjects:
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Base Sequence
Binding Sites
Clavulanic acid
Consensus Sequence
Escherichia coli
Gene Expression Regulation, Bacterial
Molecular Sequence Data
Phosphates - metabolism
Promoter Regions, Genetic
Protein Binding
Regulon
Streptomyces - chemistry
Streptomyces - genetics
Streptomyces - metabolism
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Summary:Negative control of clavulanic acid by phosphate in Streptomyces clavuligerus DSM 738 suggests that a pho regulon may exist in this bacterium. S. clavuligerus PhoP was expressed with a C-terminal His-tag in Escherichia coli and purified. Binding of PhoP-His 6 to promoter fragments of phoRP/phoU and pstS was demonstrated in gel retardation experiments. These fragments contained direct 11 bp repeats resembling PHO boxes. The tentative consensus sequence, GKTCRHBBNSV, was used to search other potential PhoP target genes in the genomic sequence of S. clavuligerus. In total, the putative PHO binding sequence was found in promoter regions of 31 S. clavuligerus genes. Binding of PhoP- His 6 to the PHO box present in the promoter region of the phosphate transporter gene SSCG_07547 of S. clavuligerus was demonstrated. Furthermore, it was shown by real time PCR that decreased concentrations of phosphate do affect increased expressions of genes to which PhoP binds. These findings confirm that a pho regulon exists in S. clavuligerus.
ISSN:1121-7138