Schultz M C, Reeder R H, Hahn S
Hutchinson Cancer Research Center, Seattle, Washington 98104.
Cell. 1992 May 15;69(4):697-702. doi: 10.1016/0092-8674(92)90233-3.
Transcription extracts prepared from yeast that are deficient in the TATA-binding protein (TBP or TFIID) are also impaired in specific promoter recognition by all three nuclear RNA polymerases (pol I, II, and III). Specific initiation can be rescued by the addition of purified recombinant TBP, demonstrating that pol I, II, and III all require this factor. A mutation of TBP has been identified that will function with pol I but not with pol II or III. Conversely, another mutation, which inactivates TATA element binding in vitro, will function with pol I and III promoters but is inactive for a pol II promoter. Thus, it is possible to identify TBP variants that will only function on different subsets of all nuclear promoters.
从缺乏TATA结合蛋白(TBP或TFIID)的酵母中制备的转录提取物,在所有三种核RNA聚合酶(聚合酶I、II和III)对特定启动子的识别方面也存在缺陷。通过添加纯化的重组TBP可以挽救特异性起始,这表明聚合酶I、II和III都需要这个因子。已经鉴定出一种TBP突变体,它能与聚合酶I起作用,但不能与聚合酶II或III起作用。相反,另一种在体外使TATA元件结合失活的突变体,能与聚合酶I和III的启动子起作用,但对聚合酶II启动子无活性。因此,有可能鉴定出仅在所有核启动子的不同子集上起作用的TBP变体。