Boam D S, Docherty K
Department of Medicine, University of Birmingham, Queen Elizabeth Hospital, Edgbaston, U.K.
Biochem J. 1989 Nov 15;264(1):233-9. doi: 10.1042/bj2640233.
Sequence-specific binding of proteins from an insulin-secreting cell line (RINm-5F) to the human insulin-gene 5' region were examined by gel-retardation and methylation-interference analysis. Specific binding of a nuclear factor to sites between nucleotides -210 to -217 and -77 to -84 was detected. The same binding activity was shown at an upstream site (-313 to -320) with low affinity. Studies using mutated binding-site probes delineated a sequence 5'-C(T/C)CTAATG-3' for high-affinity interactions. This binding activity was also present in another insulin-producing cell line (HIT.T15), but not in extracts from cell lines that did not express the insulin gene (HeLa, HL60). Cross-species comparisons show that this sequence element is highly conserved and may thus play an important role in the cell-specific regulation of insulin-gene transcription.
通过凝胶阻滞和甲基化干扰分析，检测了来自胰岛素分泌细胞系（RINm-5F）的蛋白质与人胰岛素基因5'区域的序列特异性结合。检测到一种核因子与核苷酸-210至-217和-77至-84之间位点的特异性结合。在一个上游位点（-313至-320）也显示出相同的结合活性，但亲和力较低。使用突变结合位点探针的研究确定了高亲和力相互作用的序列5'-C(T/C)CTAATG-3'。这种结合活性也存在于另一个胰岛素产生细胞系（HIT.T15）中，但在不表达胰岛素基因的细胞系（HeLa、HL60）提取物中不存在。跨物种比较表明，该序列元件高度保守，因此可能在胰岛素基因转录的细胞特异性调控中发挥重要作用。
