Gamble Matthew J, Erdjument-Bromage Hediye, Tempst Paul, Freedman Leonard P, Fisher Robert P
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Ave., New York, NY 10021, USA.
Mol Cell Biol. 2005 Jan;25(2):797-807. doi: 10.1128/MCB.25.2.797-807.2005.
To uncover factors required for transcription by RNA polymerase II on chromatin, we fractionated a mammalian cell nuclear extract. We identified the histone chaperone TAF-I (also known as INHAT [inhibitor of histone acetyltransferase]), which was previously proposed to repress transcription, as a potent activator of chromatin transcription responsive to the vitamin D3 receptor or to Gal4-VP16. TAF-I associates with chromatin in vitro and can substitute for the related protein NAP-1 in assembling chromatin onto cloned DNA templates in cooperation with the remodeling enzyme ATP-dependent chromatin assembly factor (ACF). The chromatin assembly and transcriptional activation functions are distinct, however, and can be dissociated temporally. Efficient transcription of chromatin assembled with TAF-I still requires the presence of TAF-I during the polymerization reaction. Conversely, TAF-I cannot stimulate transcript elongation when added after the other factors necessary for assembly of a preinitiation complex on naked DNA. Thus, TAF-I is required to facilitate transcription at a step after chromatin assembly but before transcript elongation.
为了揭示RNA聚合酶II在染色质上进行转录所需的因子,我们对哺乳动物细胞核提取物进行了分级分离。我们鉴定出组蛋白伴侣TAF-I(也称为INHAT[组蛋白乙酰转移酶抑制剂]),它之前被认为会抑制转录,而现在发现它是对维生素D3受体或Gal4-VP16有反应的染色质转录的有效激活剂。TAF-I在体外与染色质结合,并且在与重塑酶ATP依赖的染色质组装因子(ACF)协同作用时,能够替代相关蛋白NAP-1将染色质组装到克隆的DNA模板上。然而,染色质组装和转录激活功能是不同的,并且在时间上可以分离。用TAF-I组装的染色质的有效转录在聚合反应过程中仍然需要TAF-I的存在。相反,当在裸DNA上组装预起始复合物所需的其他因子之后添加TAF-I时,它不能刺激转录延伸。因此,在染色质组装之后但在转录延伸之前的一个步骤中,需要TAF-I来促进转录。