TFIIE orchestrates the recruitment of the TFIIH kinase module at promoter before release during transcription

Emmanuel Compe,Cathy Braun,Jean-Marc Egly,Carlos M Genes,Frederic Coin

doi:10.1038/s41467-019-10131-1

Abstract

In eukaryotes, the general transcription factors TFIIE and TFIIH assemble at the transcription start site with RNA Polymerase II. However, the mechanism by which these transcription factors incorporate the preinitiation complex and coordinate their action during RNA polymerase II transcription remains elusive. Here we show that the TFIIEα and TFIIEβ subunits anchor the TFIIH kinase module (CAK) within the preinitiation complex. In addition, we show that while RNA polymerase II phosphorylation and DNA opening occur, CAK and TFIIEα are released from the promoter. This dissociation is impeded by either ATP-γS or CDK7 inhibitor THZ1, but still occurs when XPB activity is abrogated. Finally, we show that the Core-TFIIH and TFIIEβ are subsequently removed, while elongation factors such as DSIF are recruited. Remarkably, these early transcriptional events are affected by TFIIE and TFIIH mutations associated with the developmental disorder, trichothiodystrophy.

Highlights

In eukaryotes, the general transcription factors TFIIE and TFIIH assemble at the transcription start site with RNA Polymerase II
Knowing that TTD cells with mutations in either XPB, XPD, or p8/TTD-A exhibit a decrease in the cellular concentration of TFIIH28,29, we first examined the levels of components of the transcription machinery in fibroblasts isolated from TTD patients bearing either TFIIEβ/A150P or /D187Y mutations
Such deficiency was only observed for TFIIE, since no reduction of the cellular concentration of TFIIH or for the other general transcription factors (GTFs), namely TFIIA (p35), TFIIB (p33), TFIID (TAF1, TAF4, and TBP), TFIIF (RAP74 and RAP30), and Polymerase II (Pol II) was observed

Summary

Introduction

The general transcription factors TFIIE and TFIIH assemble at the transcription start site with RNA Polymerase II. We show that while RNA polymerase II phosphorylation and DNA opening occur, CAK and TFIIEα are released from the promoter. This dissociation is impeded by either ATP-γS or CDK7 inhibitor THZ1, but still occurs when XPB activity is abrogated. We show that the Core-TFIIH and TFIIEβ are subsequently removed, while elongation factors such as DSIF are recruited These early transcriptional events are affected by TFIIE and TFIIH mutations associated with the developmental disorder, trichothiodystrophy. The general transcription factors (GTFs), including TFIIA, TFIIB, TFIID, TFIIE, TFIIF and TFIIH assemble at the transcription start site with RNA Polymerase II (Pol II). TFIIH, which intervenes during the nucleotide excision repair (NER) pathway, contains 10 subunits that can be resolved into two sub-complexes: the Core

Methods

Results

Conclusion