SITE-SPECIFIC DNA-BINDING OF NUCLEAR FACTOR-I - EFFECT OF THE SPACER REGION
SITE-SPECIFIC DNA-BINDING OF NUCLEAR FACTOR-I - EFFECT OF THE SPACER REGION
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DOI：
10.1093/nar/15.14.5545
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发表时间：
1987-07-24
影响因子：
14.9
通讯作者：
GRONOSTAJSKI, RM
中科院分区：
文献类型：
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作者：
GRONOSTAJSKI, RM
核因子 I (NFI) 是 2 型腺病毒 DNA 体外和体内复制所需的位点特异性 DNA 结合蛋白。为了研究 NFI 与 DNA 相互作用的序列要求，我们测量了蛋白质与各种合成位点的结合。 NFI 的结合位点（FIB 位点）先前显示包含由 2 个基序组成的共有序列：TGG（基序 1）和 GCCAA（基序 2），由 6 或 7bp 间隔区分隔。为了评估影响 NFI 结合的间隔区和侧翼序列中的保守序列，我们从寡核苷酸文库中分离出克隆，该寡核苷酸文库包含侧翼有 3 个简并核苷酸的两个基序，并被 6 或 7 个核苷酸的简并间隔区分开。对于 6bp 间隔区，间隔区第一个位置的 C 或 A 残基存在强偏差。具有 7bp 间隔区的位点分别在间隔区的第一个和第二个位置包含 G 和 C 或 A 残基，但在该位点的其他位置也拥有保守残基。
Nuclear factor I (NFI) is a site-specific DNA binding protein required for the replication of adenovirus type 2 DNAin vitroandin vivo. To study sequence requirements for the interaction of NFI with DNA, we have measured the binding of the protein to a variety of synthetic sites. Binding sites for NFI (FIB Sites) were previously shown to Contain a consensus sequence composed of 2 motifs, TGG (Motif 1), and GCCAA (Motif 2), separated by a 6 or 7bp spacer region. To assess conserved sequences in the spacer region and flanking sequences which affect NFI binding, we have isolated clones from oligonucleotide libraries that contain the two motifs flanked by 3 degenerate nucleotides and separated by degenerate spacer regions of 6 or 7 nucleotides. With a 6bp spacer region, a Strong bias exists for a C or A residue in the first position of the spacer. Sites with a 7bp spacer region contain a G and C or A residue at the first and second positions, respectively, of the spacer, but also possess conserved residues at other positions of the Site.