Fletcher C, Heintz N, Roeder R G
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Cell. 1987 Dec 4;51(5):773-81. doi: 10.1016/0092-8674(87)90100-0.
An octamer-binding transcription factor, OTF-1, which stimulates transcription of a human histone H2b gene, has been purified from HeLa nuclear extracts. This purification was achieved through the use of DNA affinity chromatography, and the factor was unambiguously identified by renaturation of activity following SDS-polyacrylamide gel electrophoresis. The purified factor retained the ability to efficiently stimulate H2b transcription in a reconstituted in vitro system. This effect was dependent upon an intact octamer element and was observed in the absence of the other H2b promoter elements (except the TATA motif). Furthermore, this activity was not detected in nuclear extracts prepared from cells synchronized in G2, in agreement with the in vivo data showing S-phase-specific utilization of the octamer element. From these data, we conclude that we have purified the bona fide H2b transcriptional regulatory factor.
一种能刺激人类组蛋白H2b基因转录的八聚体结合转录因子OTF-1已从HeLa细胞核提取物中纯化出来。这种纯化是通过DNA亲和层析实现的，并且在SDS-聚丙烯酰胺凝胶电泳后通过活性复性明确鉴定了该因子。纯化后的因子在重组体外系统中仍保留有效刺激H2b转录的能力。这种效应依赖于完整的八聚体元件，并且在没有其他H2b启动子元件（除TATA基序外）的情况下也能观察到。此外，在从G2期同步化的细胞制备的核提取物中未检测到这种活性，这与体内数据显示八聚体元件在S期特异性利用一致。从这些数据中，我们得出结论，我们已经纯化出了真正的H2b转录调节因子。
