Clos J, Normann A, Ohrlein A, Grummt I
Nucleic Acids Res. 1986 Oct 10;14(19):7581-95. doi: 10.1093/nar/14.19.7581.
We have determined the sequences constituting the minimal promoter of mouse rDNA. A very small region immediately upstream of the transcription start site (from -1 to -39) is sufficient to direct correct transcription initiation. Sequences immediately downstream of the transcription start site (+1 to +11) increase the efficiency of transcription initiation. Point mutations within the core promoter have been generated and assayed for their effects on template activity and on interaction with the pol I specific transcription factor TIF-IB. The core promoter element appears to consist of two functionally different domains. The distal sequence motif from position -22 to -16 is recognized by factor TIF-IB. Mutations within this region lead to similar changes of both template activity and binding of TIF-IB. Two point mutations within the proximal sequence motif from -15 to -1 do not affect TIF-IB binding although they severely impair transcription initiation. It is suggested, that this proximal region plays a role in the assembly of functional transcription initiation complexes rather than in the primary binding of TIF-IB.
我们已经确定了构成小鼠核糖体DNA最小启动子的序列。转录起始位点上游紧邻的一个非常小的区域(从-1到-39)足以指导正确的转录起始。转录起始位点下游紧邻的序列(+1到+11)可提高转录起始效率。已在核心启动子内产生点突变,并检测其对模板活性以及与RNA聚合酶I特异性转录因子TIF-IB相互作用的影响。核心启动子元件似乎由两个功能不同的结构域组成。从-22到-16的远端序列基序可被因子TIF-IB识别。该区域内的突变导致模板活性和TIF-IB结合的类似变化。从-15到-1的近端序列基序内的两个点突变虽严重损害转录起始,但不影响TIF-IB结合。有人提出,该近端区域在功能性转录起始复合物的组装中起作用,而非在TIF-IB的初级结合中起作用。