TY - JOUR TI - Requirements for RNA polymerase II preinitiation complex formation in vivo AU - Petrenko, Natalia AU - Jin, Yi AU - Dong, Liguo AU - Wong, Koon Ho AU - Struhl, Kevin A2 - Green, Michael R A2 - Manley, James L VL - 8 PY - 2019 DA - 2019/01/25 SP - e43654 C1 - eLife 2019;8:e43654 DO - 10.7554/eLife.43654 UR - https://doi.org/10.7554/eLife.43654 AB - Transcription by RNA polymerase II requires assembly of a preinitiation complex (PIC) composed of general transcription factors (GTFs) bound at the promoter. In vitro, some GTFs are essential for transcription, whereas others are not required under certain conditions. PICs are stable in the absence of nucleotide triphosphates, and subsets of GTFs can form partial PICs. By depleting individual GTFs in yeast cells, we show that all GTFs are essential for TBP binding and transcription, suggesting that partial PICs do not exist at appreciable levels in vivo. Depletion of FACT, a histone chaperone that travels with elongating Pol II, strongly reduces PIC formation and transcription. In contrast, TBP-associated factors (TAFs) contribute to transcription of most genes, but TAF-independent transcription occurs at substantial levels, preferentially at promoters containing TATA elements. PICs are absent in cells deprived of uracil, and presumably UTP, suggesting that transcriptionally inactive PICs are removed from promoters in vivo. KW - transcription KW - RNA polymerase II KW - general transcription factors KW - preinitiation complex KW - TBP-associated factors (TAFs) KW - TATA-binding protein JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -