PURIFICATION AND CHARACTERIZATION OF OTF-1, A TRANSCRIPTION FACTOR REGULATING CELL-CYCLE EXPRESSION OF A HUMAN HISTONE H2B GENE
PURIFICATION AND CHARACTERIZATION OF OTF-1, A TRANSCRIPTION FACTOR REGULATING CELL-CYCLE EXPRESSION OF A HUMAN HISTONE H2B GENE
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DOI：
10.1016/0092-8674(87)90100-0
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发表时间：
1987-12-04
期刊：
影响因子：
64.5
通讯作者：
ROEDER, RG
中科院分区：
文献类型：
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作者：
FLETCHER, C;HEINTZ, N;ROEDER, RG
八聚体结合转录因子 OTF-1 可刺激人组蛋白 H2b 基因的转录，已从 HeLa 核提取物中纯化出来。这种纯化是通过使用 DNA 亲和层析实现的，并且通过 SDS 聚丙烯酰胺凝胶电泳后的活性复性明确地鉴定了该因子。纯化的因子保留了在重建的体外系统中有效刺激 H2b 转录的能力。这种效应依赖于完整的八聚体元件，并且在不存在其他 H23b 启动子元件（TATA 基序除外）的情况下观察到。此外，在 G2 同步细胞制备的核提取物中未检测到这种活性，这与显示八聚体元素的 S 期特异性利用的体内数据一致。从这些数据中，我们得出结论，我们已经纯化了真正的 H2b 转录调节因子。
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 H23b 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 concluded that we have purified the bonafide H2b transcriptional regulatory factor.