Orphanides G, LeRoy G, Chang C H, Luse D S, Reinberg D
Howard Hughes Medical Institute, Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854, USA.
Cell. 1998 Jan 9;92(1):105-16. doi: 10.1016/s0092-8674(00)80903-4.
The requirements for transcriptional activation by RNA polymerase II were examined using chromatin templates assembled in vitro and a transcription system composed of the human general transcription factors and RNA polymerase II. Activator-induced, energy-dependent chromatin remodeling promoted efficient preinitiation complex formation and transcription initiation, but was not sufficient for productive transcription. Polymerases that initiated transcription on remodeled chromatin templates encountered a block to transcription proximal to the promoter. Entry into productive transcription required an accessory factor present in HeLa cell nuclear extract, FACT (facilitates chromatin transcription), which we have purified. FACT acts subsequent to transcription initiation to release RNA polymerase II from a nucleosome-induced block to productive transcription. The biochemical properties and polypeptide composition of FACT suggest that it is a novel protein factor that facilitates transcript elongation through nucleosomes.
利用体外组装的染色质模板以及由人类通用转录因子和RNA聚合酶II组成的转录系统,研究了RNA聚合酶II进行转录激活的要求。激活剂诱导的、能量依赖的染色质重塑促进了高效的起始前复合物形成和转录起始,但不足以进行有效的转录。在重塑的染色质模板上起始转录的聚合酶在启动子近端遇到转录障碍。进入有效的转录需要HeLa细胞核提取物中存在的一种辅助因子,即FACT(促进染色质转录),我们已将其纯化。FACT在转录起始后发挥作用,将RNA聚合酶II从核小体诱导的有效转录障碍中释放出来。FACT的生化特性和多肽组成表明它是一种新型蛋白质因子,可促进转录本通过核小体进行延伸。