Isolation and functional characterization of TIP-IB, a factor that confers promoter specificity to mouse RNA polymerase I

Andreas Schnapp, Joachim Clos, Wolfgang Hädelt, Ralf Schreck, Ales Cvekl, Ingrid Grummt

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

The murine ribosomal gene promoter contains two cis-acting control elements which operate in concert to promote efficient and accurate transcription initiation by RNA polymerase I. The start site proximal core element which is indispensable for promoter recognition by RNA polymerase I (pol I) encompasses sequences from position -39 to -1. An upstream control element (UCE) which is located between nucleotides -142 and -112 stimulates the efficiency of transcription initiation both in vivo and in vitro. Here we report the isolation and functional characterization of a specific rDNA binding protein, the transcription initiation factor TIP-IB, which specifically interacts with the core region of the mouse ribosomal RNA gene promoter. Highly purified TIP-IB complements transcriptional activity in the presence of two other essential initiation factors TIP-IA and TIF-IC. We demonstrate that the binding efficiency of purified TIF-IB to the core promoter is strongly enhanced by the presence in cis of the UCE. This positive effect of upstream sequences on TIF-IB binding is observed throughout the purification procedure suggesting that the synergistic action of the two distant promoter elements is not mediated by a protein different from TIF-IB. Increasing the distance between both control elements still facilitates stable factor binding but eliminates transcriptional activation. The results demonstrate that TIF-IB binding to the rDNA promoter is an essential early step in the assembly of a functional transcription initiation complex. The subsequent interaction of TIF-IB with other auxiliary transcription initiation factors, however, requires the correct spacing between the UCE and the core promoter element.

Original languageEnglish (US)
Pages (from-to)1385-1393
Number of pages9
JournalNucleic Acids Research
Volume18
Issue number6
StatePublished - Mar 25 1990
Externally publishedYes

Fingerprint

RNA Polymerase I
Peptide Initiation Factors
Transcription
RNA
Promoter
Isolation
Specificity
Mouse
Ribosomal DNA
Transcription Factors
rRNA Genes
Transcriptional Activation
Genes
Carrier Proteins
Nucleotides
Ribosomal RNA
Gene
Protein
Purification
Chemical activation

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Isolation and functional characterization of TIP-IB, a factor that confers promoter specificity to mouse RNA polymerase I. / Schnapp, Andreas; Clos, Joachim; Hädelt, Wolfgang; Schreck, Ralf; Cvekl, Ales; Grummt, Ingrid.

In: Nucleic Acids Research, Vol. 18, No. 6, 25.03.1990, p. 1385-1393.

Research output: Contribution to journalArticle

Schnapp, Andreas ; Clos, Joachim ; Hädelt, Wolfgang ; Schreck, Ralf ; Cvekl, Ales ; Grummt, Ingrid. / Isolation and functional characterization of TIP-IB, a factor that confers promoter specificity to mouse RNA polymerase I. In: Nucleic Acids Research. 1990 ; Vol. 18, No. 6. pp. 1385-1393.
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