TAF(II)-independent activation mediated by human TBP in the presence of the positive cofactor PC4

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

TFIID is a multiprotein complex comprised of the TATA-binding protein (TBP) and an array of TBP-associated factors (TAF(II)s). Whereas TBP is sufficient for basal transcription in conjunction with other general transcription factors and RNA polymerase II, TAF(II)s are additionally required for activator-dependent transcription in mammalian cell-free transcription systems. However, recent in vivo studies carried out in yeast suggest that TAF(II)s are not globally required for activator function. The discrepancy between in vivo yeast studies and in vitro mammalian cell-free systems remains to be resolved. In this study, we describe a mammalian cell-free transcription system reconstituted with only recombinant proteins and epitope-tagged multiprotein complexes. Transcriptional activation can be recapitulated in this highly purified in vitro transcription system in the absence of TAF(II)s. This TBP-mediated activation is not induced by human mediator, another transcriptional coactivator complex potentially implicated in activator response. In contrast, general transcription factors TFIIH and TFIIA play a significant role in TBP-mediated activation, which can be detected in vitro with Gal4 fusion proteins containing various transcriptional activation domains. Our data, therefore, suggest that TFIIH and TFIIA can mediate activator function in the absence of TAF(II)s.

Original languageEnglish (US)
Pages (from-to)4478-4490
Number of pages13
JournalEMBO Journal
Volume17
Issue number15
DOIs
StatePublished - Aug 3 1998

Fingerprint

TATA-Box Binding Protein
Transcription
Cell-Free System
Transcription Factor TFIIA
Chemical activation
General Transcription Factors
Activator Appliances
Multiprotein Complexes
Transcriptional Activation
Cells
Transcription Factor TFIIH
TATA-Binding Protein Associated Factors
Yeasts
Yeast
Transcription Factor TFIID
Protein Array Analysis
RNA Polymerase II
Recombinant Proteins
Epitopes
Fusion reactions

Keywords

  • TAF(II)s
  • TBP
  • TFIIA
  • TFIIH
  • Transcriptional activation

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

TAF(II)-independent activation mediated by human TBP in the presence of the positive cofactor PC4. / Wu, Shwu Yuan; Kershnar, Eric; Chiang, Cheng Ming.

In: EMBO Journal, Vol. 17, No. 15, 03.08.1998, p. 4478-4490.

Research output: Contribution to journalArticle

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