TFIIA and the transactivator Rap1 cooperate to commit TFIID for transcription initiation

Transcription of eukaryotic mRNA encoding genes by RNA polymerase II (Pol II) is triggered by the binding of transactivating proteins to enhancer DNA, which stimulates the recruitment of general transcription factors (GTFs; TFIIA, B, D, E, F, H) and Pol II on the cis-linked promoter leading to preinitiation complex (PIC) formation and transcription1. In TFIID-dependent activation pathways, this TATA box Binding Protein (TBP)-containing GTF is first recruited on the promoter through interaction with activators1-3 and cooperates with TFIIA to form a committed PIC4. However, neither the mechanisms by which activation signals are communicated between these factors, nor the structural organization of the activated PIC are known. Here we used cryo-electron microscopy to determine the architecture of nucleoprotein complexes composed of TFIID, TFIIA, the transcriptional activator Rap1 and yeast enhancer-promoter DNA. These structures revealed the mode of binding of Rap1 and TFIIA to TFIID, as well as a reorganization of TFIIA induced by its interaction with Rap1. We propose that this change in position increases the exposure of TBP within TFIID, consequently enhancing its ability to interact with the promoter. A large Rap1-dependent DNA loop forms between the activator binding site and the proximal promoter region, and this loop is topologically locked by a TFIIA-Rap1 protein bridge that folds over the DNA. These results highlight the role of TFIIA in transcriptional activation, define a molecular mechanism for enhancer-promoter communication and provide important new structural insights into the pathways of intramolecular communication that convey transcription activation signals through the TFIID complex.