Reconstitution of yeast RNA polymerase I transcription in vitro from purified components. TATA-binding protein is not required for basal transcription

J Biol Chem. 1998 Dec 11;273(50):33795-802. doi: 10.1074/jbc.273.50.33795.

Abstract

Five purified protein components, RNA polymerase I, Rrn3p, core factor, TBP (TATA-binding protein), and upstream activation factor, are sufficient for high level transcription in vitro from the Saccharomyces cerevisiae rDNA promoter. Rrn3p and pol I form a complex in solution that is active in specific initiation. Three protein components, pol I, Rrn3p, and core factor, and promoter sequence to -38, suffice for basal transcription. Unlike pol II and pol III, yeast pol I basal transcription does not require TBP. Instead, TBP, upstream activation factor, and the upstream element of the promoter together stimulate pol I basal transcription to a fully activated level. The role of TBP in pol I transcription is fundamentally different from its role in pol II or pol III transcription.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • DNA, Ribosomal / genetics
  • DNA-Binding Proteins / metabolism*
  • Fungal Proteins / metabolism
  • Molecular Sequence Data
  • Pol1 Transcription Initiation Complex Proteins*
  • Promoter Regions, Genetic
  • RNA Polymerase I / genetics*
  • RNA Polymerase I / metabolism
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae Proteins*
  • TATA Box
  • TATA-Box Binding Protein
  • Transcription Factors / metabolism*
  • Transcription, Genetic*

Substances

  • DNA, Ribosomal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Pol1 Transcription Initiation Complex Proteins
  • RRN3 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • TATA-Box Binding Protein
  • Transcription Factors
  • RNA Polymerase I