Reconstitution of transcription with five purified initiation factors and RNA polymerase II from Saccharomyces cerevisiae.
Reconstitution of transcription with five purified initiation factors and RNA polymerase II from Saccharomyces cerevisiae.
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DOI：
10.1016/s0021-9258(18)50101-0
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发表时间：
1992-11
期刊：
影响因子：
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通讯作者：
M. H. Sayre;Herbert Tschochner;R. Kornberg
中科院分区：
文献类型：
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作者：
M. H. Sayre;Herbert Tschochner;R. Kornberg
从全细胞提取物中分离出酵母RNA聚合酶II起始因子a、B、e和g，发现当与细菌表达的酵母转录因子（TF）IID结合时，足以使RNA聚合酶II在体外利用9种不同的真核启动子，并在体内使用的位点起始转录。纯化的因子不含先前描述的转录因子IIA（TFIIA）。TFIIA不能替代任何纯化的因子或用全套因子刺激转录，表明其在粗提物中的功能主要是作为抗抑制剂。
Yeast RNA polymerase II initiation factors a, b, e, and g were isolated from whole cell extract and found to be sufficient, when combined with bacterially expressed yeast transcription factor (TF) IID, to enable RNA polymerase II to utilize nine different eukaryotic promoters in vitro, and to initiate transcription at sites used in vivo. The purified factors did not contain the previously described transcription factor IIA (TFIIA). TFIIA failed to substitute for any purified factor or to stimulate transcription with the complete set of factors, indicating that its function in crude extracts is primarily as an anti-inhibitor.