Mason Paul B, Struhl Kevin
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Mol Cell Biol. 2003 Nov;23(22):8323-33. doi: 10.1128/MCB.23.22.8323-8333.2003.
The FACT complex facilitates transcription on chromatin templates in vitro, and it has been functionally linked to nucleosomes and putative RNA polymerase II (Pol II) elongation factors. In Saccharomyces cerevisiae cells, FACT specifically associates with active Pol II genes in a TFIIH-dependent manner and travels across the gene with elongating Pol II. Conditional inactivation of the FACT subunit Spt16 results in increased Pol II density, transcription, and TATA-binding protein (TBP) occupancy in the 3' portion of certain coding regions, indicating that FACT suppresses inappropriate initiation from cryptic promoters within coding regions. Conversely, loss of Spt16 activity reduces the association of TBP, TFIIB, and Pol II with normal promoters. Thus, FACT is required for wild-type cells to restrict initiation to normal promoters, thereby ensuring that only appropriate mRNAs are synthesized. We suggest that FACT contributes to the fidelity of Pol II transcription by linking the processes of initiation and elongation.
FACT复合物在体外促进染色质模板上的转录,并且在功能上与核小体和假定的RNA聚合酶II(Pol II)延伸因子相关联。在酿酒酵母细胞中,FACT以TFIIH依赖的方式特异性地与活跃的Pol II基因相关联,并随着延伸的Pol II穿越基因。FACT亚基Spt16的条件性失活导致某些编码区3'部分的Pol II密度、转录和TATA结合蛋白(TBP)占有率增加,这表明FACT抑制了编码区内隐蔽启动子的不适当起始。相反,Spt16活性的丧失会降低TBP、TFIIB和Pol II与正常启动子的关联。因此,野生型细胞需要FACT将起始限制在正常启动子上,从而确保只合成合适的mRNA。我们认为FACT通过连接起始和延伸过程来促进Pol II转录的保真度。