Expression and heat-responsive regulation of a TFIIB homologue from the archaeon Haloferax volcanii

Mol Microbiol. 1999 Sep;33(5):1081-92. doi: 10.1046/j.1365-2958.1999.01551.x.

Abstract

Multiple divergent genes encoding the eukaryal-like TFIIB (TFB) transcription initiation factor have been identified in the archaeon Haloferax volcanii. Expression of one of these TFB-encoding genes, referred to here as tfb2, was induced specifically in response to heat shock at the transcription level. A time course for tfb2 induction demonstrated that mRNA levels increased as much as eightfold after 15 min at 60 degrees C. A transcription fusion of the tfb2 promoter region with a stable RNA reporter gene confirmed the heat responsiveness of the tfb2 core promoter, and immunoblot analysis using antibodies generated against a recombinant His-tagged TFB2 showed that the protein levels of one TFB increased slightly in response to elevated temperatures. An archaeal consensus TATA element (5'-TTTATA-3') was located 110 bp upstream of the translation start site and appeared to be used for both basal and heat shock-induced expression. The long DNA leader region (79 bp) preceding the predicted AUG translation start codon for TFB2 contained a T-rich sequence element located 22 bp downstream of the transcription start site. Using an in vivo transcription termination assay, we demonstrated that this T-rich element can function as a sequence-dependent transcription terminator, which may serve to downregulate expression of the tfb2 gene under both non-heat shock and heat shock conditions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Archaeal Proteins / genetics*
  • Archaeal Proteins / metabolism
  • Base Sequence
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Gene Expression Regulation, Archaeal*
  • Genes, Reporter
  • Haloferax volcanii / genetics*
  • Haloferax volcanii / physiology
  • Heat-Shock Response
  • Hot Temperature
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Regulatory Sequences, Nucleic Acid
  • Transcription Factor TFIIB
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • Archaeal Proteins
  • Recombinant Proteins
  • Transcription Factor TFIIB
  • Transcription Factors

Associated data

  • GENBANK/AF143963