Gronostajski R M, Knox J, Berry D, Miyamoto N G
Ontario Cancer Institute, University of Toronto, Canada.
Nucleic Acids Res. 1988 Mar 25;16(5):2087-98. doi: 10.1093/nar/16.5.2087.
Nuclear factor I (NFI) is a site-specific DNA binding protein required for the replication of adenovirus DNA in vitro and in vivo. We have examined the effect of natural and synthetic binding sites for NFI (FIB sites) on RNA synthesis in HeLa whole cell extracts. The natural binding site used is the 26bp FIB-2 site previously isolated from the human genome. When present upstream of the TATA box of the adenovirus major late promoter, the FIB-2 site stimulates RNA synthesis 3 to 5-fold. This stimulation occurs with either orientation of the FIB-2 site. A point mutation in FIB-2 that decreases NFI binding at least 100-fold reduces, but does not completely abolish, the stimulation of transcription. A number of synthetic binding sites for NFI were tested for the ability to increase RNA synthesis. The strongest binding sites stimulated transcription the most, while the weakest sites had the least effect. These studies strongly suggest a role for NFI and cellular FIB sites in the control of RNA synthesis.
核因子I（NFI）是一种位点特异性DNA结合蛋白，在体外和体内腺病毒DNA复制过程中都是必需的。我们已经研究了NFI的天然和合成结合位点（FIB位点）对HeLa全细胞提取物中RNA合成的影响。所使用的天然结合位点是先前从人类基因组中分离出的26bp FIB-2位点。当位于腺病毒主要晚期启动子的TATA框上游时，FIB-2位点可刺激RNA合成3至5倍。无论FIB-2位点的方向如何，这种刺激都会发生。FIB-2中的一个点突变使NFI结合减少至少100倍，该突变会降低但不会完全消除对转录的刺激。测试了许多NFI的合成结合位点增加RNA合成的能力。结合能力最强的位点对转录的刺激作用最大，而结合能力最弱的位点影响最小。这些研究有力地表明了NFI和细胞FIB位点在RNA合成控制中的作用。
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