The archaeal TFIIEalpha homologue facilitates transcription initiation by enhancing TATA-box recognition

EMBO Rep. 2001 Feb;2(2):133-8. doi: 10.1093/embo-reports/kve021.

Abstract

Transcription from many archaeal promoters can be reconstituted in vitro using recombinant TATA-box binding protein (TBP) and transcription factor B (TFB)--homologues of eukaryal TBP and TFIIB--together with purified RNA polymerase (RNAP). However, all archaeal genomes sequenced to date reveal the presence of TFE, a homologue of the alpha-subunit of the eukaryal general transcription factor, TFIIE. We show that, while TFE is not absolutely required for transcription in the reconstituted in vitro system, it nonetheless plays a stimulatory role on some promoters and under certain conditions. Mutagenesis of the TATA box or reduction of TBP concentration in transcription reactions sensitizes a promoter to TFE addition. Conversely, saturating reactions with TBP de-sensitizes promoters to TFE. These results suggest that TFE facilitates or stabilizes interactions between TBP and the TATA box.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / metabolism*
  • Base Sequence
  • DNA Footprinting
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed RNA Polymerases / metabolism
  • Deoxyribonuclease I / metabolism
  • Molecular Sequence Data
  • Mutation / genetics
  • Protein Binding
  • Protein Subunits
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Sulfolobus / genetics*
  • TATA Box / genetics*
  • TATA-Box Binding Protein
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*
  • Transcription Factors, TFII*
  • Transcription, Genetic*

Substances

  • Archaeal Proteins
  • DNA-Binding Proteins
  • Protein Subunits
  • TATA-Box Binding Protein
  • Transcription Factors
  • Transcription Factors, TFII
  • transcription factor TFIIE
  • DNA-Directed RNA Polymerases
  • Deoxyribonuclease I