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PCR Cloning and Subcloning> | … Coding sequences for Rrn5, Rrn9, Rrn10, and Uaf30 were codon optimized, synthesized (Genscript), and subcloned. Autoinduction was carried out in LOBSTR-BL21(DE3) cells (… | Get A Quote |
Preribosomal RNA is selectively transcribed by RNA polymerase (Pol) I in eukaryotes. The yeast transcription factor upstream activating factor (UAF) represses Pol II transcription and mediates Pol I preinitiation complex (PIC) formation at the 35 ribosomal RNA gene. To visualize the molecular intermediates toward PIC formation, we determined the structure of UAF in complex with native promoter DNA and transcription factor TATA-box-binding protein (TBP). We found that UAF recognizes DNA using a hexameric histone-like scaffold with markedly different interactions compared with the nucleosome and the histone-fold-rich transcription factor IID (TFIID). In parallel, UAF positions TBP for Core Factor binding, which l... More