Autogenous transcriptional activation of a thiostrepton-induced gene in Streptomyces lividans

EMBO J. 1993 Aug;12(8):3183-91.

Abstract

Although the antibiotic thiostrepton is best known as an inhibitor of protein synthesis, it also, at extremely low concentrations (< 10(-9) M), induces the expression of a regulon of unknown function in certain Streptomyces species. Here, we report the purification of a Streptomyces lividans thiostrepton-induced transcriptional activator protein, TipAL, whose N-terminus is similar to a family of eubacterial regulatory proteins represented by MerR. TipAL was first purified from induced cultures of S.lividans as a factor which bound to and activated transcription from its own promoter. The tipAL gene was overexpressed in Escherichia coli and TipAL protein purified in a single step using a thiostrepton affinity column. Thiostrepton enhanced binding of TipAL to the promoter and catalysed specific transcription in vitro. TipAS, a second gene product of the same open reading frame consisting of the C-terminal domain of TipAL, is apparently translated using its own in-frame initiation site. Since it is produced in large molar excess relative to TipAL after induction and also binds thiostrepton, it may competitively modulate transcriptional activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Bacterial Proteins / metabolism
  • Base Sequence
  • DNA, Bacterial / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / metabolism
  • Gene Expression Regulation, Bacterial* / drug effects
  • Molecular Sequence Data
  • Open Reading Frames
  • Protein Binding
  • RNA, Bacterial
  • Sequence Analysis
  • Streptomyces / drug effects
  • Streptomyces / genetics*
  • Thiostrepton / pharmacology*
  • Trans-Activators / genetics
  • Trans-Activators / isolation & purification*
  • Trans-Activators / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / isolation & purification*
  • Transcription Factors / metabolism
  • Transcription, Genetic / drug effects

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA-Binding Proteins
  • RNA, Bacterial
  • TipAL protein, Streptomyces
  • Trans-Activators
  • Transcription Factors
  • tipAS protein, Streptomyces
  • Thiostrepton