The architecture of human general transcription factor TFIID core complex

Abstract

General transcription factor TFIID is a key component of RNA polymerase II transcription initiation. Human TFIID is a megadalton-sized complex comprising TATA-binding protein (TBP) and 13 TBP-associated factors (TAFs). TBP binds to core promoter DNA, recognizing the TATA-box. We identified a ternary complex formed by TBP and the histone fold (HF) domain-containing TFIID subunits TAF11 and TAF13. We demonstrate that TAF11/TAF13 competes for TBP binding with TATA-box DNA, and also with the N-terminal domain of TAF1 previously implicated in TATA-box mimicry. In an integrative approach combining crystal coordinates, biochemical analyses and data from cross-linking mass-spectrometry (CLMS), we determine the architecture of the TAF11/TAF13/TBP complex, revealing TAF11/TAF13 interaction with the DNA binding surface of TBP. We identify a highly conserved C-terminal TBP-interaction domain (CTID) in TAF13, which is essential for supporting cell growth. Our results thus have implications for cellular TFIID assembly and suggest a novel regulatory state for TFIID function.

Structural and functional insight into TAF1–TAF7, a subcomplex of transcription factor II D

“…TFIID nucleates the formation of the preinitiation complex (PIC) at the transcriptional start site by recruiting other general transcription factors (TFII-A, -B, -E, -F, and -H), along with RNA Pol II. TFIID is assembled in a stepwise manner, with a symmetric TFIID core complex recruited first, followed by further recruitment of additional TATA binding protein (TBP)-associated factors (TAFs) to form the complete asymmetric holo-TFIID complex (1). This megadalton-sized multiprotein assembly, comprised of TBP and 13 evolutionary conserved TAFs (2), is organized into a trilobed structure (3) and undergoes striking rearrangements upon binding to TFIIA and DNA (4).…”

A variant inTAF1is associated with a new syndrome with severe intellectual disability and characteristic dysmorphic features

“…We were able to prioritize variants that were not observed in other members of the sequenced family, including not being present in an unaffected male cousin. The only variant found in both of our two variant prioritization schemes is in a highly conserved region of TAF1, which is the largest subunit of the TFIID multi-protein complex involved in transcription initiation [49,[54][55][56][57].…”

The MultiBac Baculovirus/Insect Cell Expression Vector System for Producing Complex Protein Biologics

“…MultiBac produced material for electron microscopic studies include the structures of COP1-coated vesicles, revealing alternate coatomer conformations and interactions [66,67], the architecture of the physiological core of human general transcription factor TFIID [52,58], or the elucidation of the molecular mechanisms of the anaphase promoting complex APC/C at sub-nanometer resolution [68]. Recently, the entire human Mediator transcription factor holocomplex has been successfully assembled by using MultiBac, and functionally characterized [69].…”

“…In contrast to very late promoters such as polh and p10, earlier promoters are expressed at lower levels. However, it may be often impractical to resort to co-infection or to tuning protein expression levels by promoter choice, also due to the fact that the timing of the production of protein subunits will then be altered as highly successful in producing difficult-to-express protein complexes in high quality and quantity, including the physiological core complex of human general transcription factor TFIID [52][53][54]. A notable exploit is influenza polymerase, an important drug target to combat the flu, which has remained inaccessible for 40 years since its discovery.…”