A role for ATP and TFIIH in activation of the RNA polymerase II preinitiation complex prior to transcription initiation

Arik Dvir, Karla Pfeil Garrett, Christian Chalut, Jean Marc Egly, Joan Weliky Conaway, Ronald C. Conaway

Research output: Contribution to journalArticlepeer-review

72 Scopus citations

Abstract

A requirement for an ATP cofactor in synthesis of the first 8-10 bonds of promoter-specific transcripts by RNA polymerase II is well established. Whether ATP is required for synthesis of the first phosphodiester bond or at a slightly later stage in synthesis of nascent transcripts, however, remains controversial. Goodrich and Tjian (Goodrich, J. A., and Tjian, R. (1994) Cell 77, 145-156) recently proposed that synthesis of the first phosphodiester bond of promoter-specific transcripts by RNA polymerase II is independent of ATP and general transcription factors TFIIE and TFIIH. Here we investigate this model. Taken together, our findings indicate that ATP, TFIIE, and TFIIH can have a profound effect on the efficiency of transcription initiation. First, we observe that synthesis of the first phosphodiester bond of transcripts initiated at the adenovirus 2 major late promoter depends strongly on ATP, TFIIE, and TFIIH in a transcription system reconstituted with RNA polymerase II, TFIIH, and recombinant TBP, TFIIB, TFIIE, and TFIIF. Second, we demonstrate that, in this enzyme system, ATP-dependent activation of transcription initiation can occur immediately prior to synthesis of the first phosphodiester bond of nascent transcripts. Finally, we demonstrate that the activated initiation complex is unstable and decays rapidly to an inactive state in the presence of the inhibitor ATPγS (adenosine 5'- O(thio)triphosphate), even during reiterative synthesis of abortive transcripts.

Original languageEnglish (US)
Pages (from-to)7245-7248
Number of pages4
JournalJournal of Biological Chemistry
Volume271
Issue number13
DOIs
StatePublished - Mar 29 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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