A TISSUE-SPECIFIC NUCLEAR FACTOR BINDS TO MULTIPLE SITES IN THE HUMAN INSULIN-GENE ENHANCER
A TISSUE-SPECIFIC NUCLEAR FACTOR BINDS TO MULTIPLE SITES IN THE HUMAN INSULIN-GENE ENHANCER
复制标题
DOI：
10.1042/bj2640233
复制
发表时间：
1989-11-15
影响因子：
4.1
通讯作者：
DOCHERTY, K
中科院分区：
文献类型：
--
作者：
BOAM, DSW;DOCHERTY, K
用凝胶滞留和甲基化干扰分析检测了胰岛素分泌细胞系(RINm-5F)的蛋白质与人胰岛素基因5‘’区的序列特异性结合。检测到核因子与核苷酸-210至-217和-77至-84之间的位点的特异性结合。在低亲和力的上游部位(-313~-320)也显示出相同的结合活性。使用突变的结合位点探针的研究描绘了5‘’-C(T/C)CTAATG-3‘’序列，用于高亲和力相互作用。这种结合活性也存在于另一种胰岛素产生细胞系(HIT.T15)中，但不存在于不表达胰岛素基因的细胞系(HeLa，HL60)的提取物中。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。
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.