Unique TATA-binding protein-containing complexes and cofactors involved in transcription by RNA polymerases II and III

Cheng Ming Chiang, Hui Ge, Zhengxin Wang, Alexander Hoffmann, Robert G. Roeder

Research output: Contribution to journalArticle

165 Citations (Scopus)

Abstract

Two multisubunit complexes containing the TATA-binding protein (TBP) were isolated from HeLa cells constitutively expressing the FLAG epitope-tagged TBP using antibody affinity and peptide elution methods. One of the complexes (f:TFIID), isolated from the P11 0.85 M KCl fraction, contains at least 13 specific TBP-associated factors (TAFs) and can mediate activator-dependent transcription by RNA polymerase II. Importantly, activator function through the highly purified f:TFIID complex still requires a general cofactor fraction containing upstream factor stimulatory activity (USA). As previously observed with partially purified activator-competent natural TFQD, f:TFIID generates extended TATA-dependent footprints on the intrinsically strong adenovirus major late promoter (MLP) but only restricted footprints on the weak adenovirus E1b and E4 and HIV (core) promoters. Along with previous demonstrations of activator-induced downstream TFIID interactions on the E4 promoter, these results argue for a relationship between downstream interactions and overall promoter strength. Initiator-like sequences appear not to be essential for downstream interactions since they have no effect on downstream MLP interactions when mutated, do not effect downstream interactions on the HIV promoter and are not present on the inducible E4 promoter. The other multisubunit complex (f:TFIIIB), isolated from the P11 0.30 M KCl fraction, contains four specific TAFs and can substitute for one of the fractions (TFIIIB) required for RNA polymerase III (pol III) transcription. Neither f:TFIID nor TBP could substitute for this pol III TBP-containing fraction. This plus the fact that f:TFIIIB failed to generate a footprint on the MLP underscores the importance of TAFs in determining promoter specificity by different RNA polymerases.

Original languageEnglish (US)
Pages (from-to)2749-2762
Number of pages14
JournalEMBO Journal
Volume12
Issue number7
StatePublished - 1993

Fingerprint

Transcription Factor TFIID
TATA-Box Binding Protein
RNA Polymerase III
RNA Polymerase II
Transcription
Transcription Factor TFIIIB
TATA-Binding Protein Associated Factors
Adenoviridae
Upstream Stimulatory Factors
HIV
Activator Appliances
Antibody Affinity
DNA-Directed RNA Polymerases
HeLa Cells
Epitopes
Demonstrations
Peptides
Antibodies

Keywords

  • Epitope-tagged TBP
  • TAF
  • TFIID
  • TFIIIB
  • Transcription

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

Unique TATA-binding protein-containing complexes and cofactors involved in transcription by RNA polymerases II and III. / Chiang, Cheng Ming; Ge, Hui; Wang, Zhengxin; Hoffmann, Alexander; Roeder, Robert G.

In: EMBO Journal, Vol. 12, No. 7, 1993, p. 2749-2762.

Research output: Contribution to journalArticle

Chiang, Cheng Ming ; Ge, Hui ; Wang, Zhengxin ; Hoffmann, Alexander ; Roeder, Robert G. / Unique TATA-binding protein-containing complexes and cofactors involved in transcription by RNA polymerases II and III. In: EMBO Journal. 1993 ; Vol. 12, No. 7. pp. 2749-2762.
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