McKeon C, Pastan I, de Crombrugghe B
Nucleic Acids Res. 1984 Apr 25;12(8):3491-502. doi: 10.1093/nar/12.8.3491.
The chromatin structure of the chick alpha 2(I) collagen gene was probed with DNase I. Because our previous work strongly suggested that the 5' end of this gene is not methylated whereas the rest of the gene is methylated whether or not the gene is expressed, we compared the relative DNase I sensitivity of the methylated and unmethylated segments. Both regions demonstrate similar relative DNase I sensitivities within a given tissue. In chromatin of chick embryo fibroblasts, we find a DNase I hypersensitive site which maps between 100 and 300 bp preceding the start of transcription. This site is not found in brain chromatin but is present in chick embryo fibroblasts transformed by Rous Sarcoma virus although the rate of transcription of the alpha 2(I) collagen gene is greatly reduced in these cells. Hence, the mechanism responsible for the large decrease in alpha 2(I) collagen gene expression in RSV transformed cells is different from the mechanism that is responsible for the presence of a DNase I hypersensitive site in the promoter. Furthermore, changes in the DNase I sensitivity of the chromatin of the alpha 2(I) collagen promoter occur without changes in the methylation pattern of the gene.
用脱氧核糖核酸酶I(DNase I)探测了鸡α2(I)型胶原基因的染色质结构。由于我们之前的研究强烈表明,该基因的5'端未发生甲基化,而无论该基因是否表达,基因的其余部分均发生甲基化,因此我们比较了甲基化和未甲基化片段对DNase I的相对敏感性。在给定组织中,这两个区域表现出相似的对DNase I的相对敏感性。在鸡胚成纤维细胞的染色质中,我们发现一个DNase I超敏位点,它位于转录起始点之前100至300 bp之间。在脑染色质中未发现该位点,但在经劳氏肉瘤病毒转化的鸡胚成纤维细胞中存在,尽管这些细胞中α2(I)型胶原基因的转录速率大大降低。因此,劳氏肉瘤病毒转化细胞中α2(I)型胶原基因表达大幅下降的机制与启动子中存在DNase I超敏位点的机制不同。此外,α2(I)型胶原启动子染色质对DNase I敏感性的变化并未伴随该基因甲基化模式的改变。