Identification of a protein-binding sequence involved in expression of an esterase gene from Streptomyces scabies

M J Babcock, M McGrew, J L Schottel

Research output: Contribution to journalArticlepeer-review

Abstract

Expression of an esterase gene from Streptomyces scabies is regulated by zinc in both Streptomyces scabies and Streptomyces lividans. A specific protein-binding site was identified on an esterase promoter fragment by using an S-30 extract from S. scabies. The location of the protein-binding site was determined by gel shift assays of promoter deletion fragments and by DNase I footprinting analysis. The protein-binding site maps from nucleotides -59 to -81 relative to the start of transcription. An esterase gene construct cloned and expressed in S. lividans was used to assess the importance of the protein-binding site. Deletion of the 23-bp protein-binding site resulted in a 10-fold decrease in esterase production when cells were grown in zinc-inducing conditions. The protein-binding site may represent a region involved in positive regulation of the S. scabies esterase gene.
Original languageEnglish
Pages (from-to)4287-93
Number of pages7
JournalJournal of Bacteriology
Volume174
Issue number13
Publication statusPublished - Jul 1992

Keywords

  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Chromosome Deletion
  • Cloning, Molecular
  • DNA, Bacterial
  • Deoxyribonuclease I
  • Esterases
  • Gene Expression Regulation, Bacterial
  • Gene Expression Regulation, Enzymologic
  • Genes, Bacterial
  • Kinetics
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Oligodeoxyribonucleotides
  • Protein Binding
  • Restriction Mapping
  • Streptomyces
  • Transcription, Genetic
  • Transformation, Bacterial

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