Satyamoorthy K, Park K, Atchison M L, Howe C C
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania.
Mol Cell Biol. 1993 Nov;13(11):6621-8. doi: 10.1128/mcb.13.11.6621-6628.1993.
Murine intracisternal A-particle long terminal repeats contain an intracisternal A-particle upstream enhancer (IUE) element that binds to a 65-kDa IUE binding protein (IUEB) present in both undifferentiated F9 embryonal carcinoma cells and differentiated parietal endoderm-like PYS-2 cells. This IUE element confers a CpG methylation-sensitive IUEB binding and enhancer activity. Using gel retardation, methylation interference, CpG methylation sensitivity binding, and cotransfection assays, we have now identified the 65-kDa IUEB as YY1 (also called NF-E1, delta, or UCRBP), a zinc finger protein related to the Krüppel family. YY1 binds to a number of similar but distinct DNA motifs, and cotransfection assays indicate that these motifs have different enhancer potentials in PYS-2 cells. The relative strengths of these elements are as follows: IUE > kappa E3' from the human immunoglobulin kappa light-chain 3' enhancer > upstream conserved region from the Moloney murine leukemia virus promoter. Results of DNA binding assays suggest that the differences in enhancer potentials are due to the different binding affinities of YY1 to the various motifs and the binding of two other transcription factors to the IUE sequence.
小鼠脑池内A颗粒长末端重复序列包含一个脑池内A颗粒上游增强子(IUE)元件,该元件可与未分化的F9胚胎癌细胞和分化的壁内胚层样PYS-2细胞中均存在的65 kDa IUE结合蛋白(IUEB)结合。这个IUE元件赋予了对CpG甲基化敏感的IUEB结合和增强子活性。通过凝胶阻滞、甲基化干扰、CpG甲基化敏感性结合和共转染分析,我们现已确定65 kDa的IUEB为YY1(也称为NF-E1、δ或UCRBP),一种与Krüppel家族相关的锌指蛋白。YY1可与许多相似但不同的DNA基序结合,共转染分析表明这些基序在PYS-2细胞中具有不同的增强子潜能。这些元件的相对强度如下:IUE>来自人免疫球蛋白κ轻链3′增强子的κE3′>来自莫洛尼鼠白血病病毒启动子的上游保守区。DNA结合分析结果表明,增强子潜能的差异是由于YY1对各种基序的不同结合亲和力以及另外两种转录因子与IUE序列的结合所致。