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Item Mediator and SAGA Have Distinct Roles in Pol II Preinitiation Complex Assembly and Function(Elsevier, 2012) Chen, Xiao-Fen; Lehmann, Lynn; Lin, Justin J.; Vashisht, Ajay; Schmidt, Ryan; Ferrari, Roberto; Huang, Chengyang; McKee, Robin; Mosley, Amber; Plath, Kathrin; Kurdistani, Siavash K.; Wohlschlegel, James; Carey, Michael; Biochemistry and Molecular Biology, School of MedicineA key feature of RNA polymerase II preinitiation complexes (PICs) is their ability to coordinate transcription initiation with chromatin modification and remodeling. To understand how this coordination is achieved, we employed extensive proteomic and mechanistic analyses to study the composition and assembly of PICs in HeLa and mouse embryonic stem (ES) cell nuclear extracts. Strikingly, most of the machinery necessary for transcription initiation on chromatin is part of the PIC. The PIC is nearly identical between ES and HeLa cells, and contains two major co-activator complexes, Mediator and SAGA. Genomewide analysis of Mediator reveals a close correlation with Pol II, TBP and mRNA levels implying a major role in PIC assembly. Moreover, Mediator coordinates assembly of the Pol II initiation factors and chromatin machinery into a PIC in vitro, while SAGA acts after PIC assembly to allow transcription on chromatin.