Salditt-Georgieff M, Sheffery M, Krauter K, Darnell J E, Rifkind R, Marks P A
J Mol Biol. 1984 Feb 5;172(4):437-50. doi: 10.1016/s0022-2836(84)80016-9.
The transcription of the beta-globin genes in mouse erythroleukemia cells has been examined by hybridizing labeled RNA obtained from isolated nuclei after chain elongation in the presence of [alpha-32P]UTP. There is induction of at least 30-fold of beta maj globin transcription after cells are treated with either dimethylsulfoxide or hexamethylene bisacetamide. The induction requires 36 to 48 hours to be maximal, during which time the cells double about three to four times. During this time, a site in the beta maj DNA region becomes hypersensitive to DNase. The development of this hypersensitive site is co-ordinate with the transcriptional increase. The induced transcripts in the beta-globin region are alpha-amanitin-sensitive (and therefore are RNA polymerase II products). An examination of weak transcriptional signals to DNA fragments upstream of the beta maj globin gene in uninduced mouse erythroleukemia cells and in cells that do not make globin is also reported. The low level of hybridization to the upstream regions in uninduced erythroleukemia cells, in L cells (a fibroblast) and in a strain of erythroleukemia cells that no longer make globin are not equally sensitive to alpha-amanitin as in the induced signal. These experiments help define the inducible transcription unit for beta maj globin mRNA production.
通过使在[α-32P]UTP存在下进行链延伸后从分离的细胞核中获得的标记RNA杂交,研究了小鼠红白血病细胞中β-珠蛋白基因的转录情况。在用二甲基亚砜或六亚甲基双乙酰胺处理细胞后,βmaj珠蛋白转录至少诱导了30倍。诱导需要36至48小时达到最大值,在此期间细胞大约倍增三到四次。在此期间,βmaj DNA区域中的一个位点对DNase变得高度敏感。这种高度敏感位点的出现与转录增加是同步的。β-珠蛋白区域中的诱导转录本对α-鹅膏蕈碱敏感(因此是RNA聚合酶II的产物)。还报道了对未诱导的小鼠红白血病细胞和不产生珠蛋白的细胞中βmaj珠蛋白基因上游DNA片段的弱转录信号的研究。未诱导的红白血病细胞、L细胞(一种成纤维细胞)和不再产生珠蛋白的红白血病细胞株中与上游区域的低水平杂交对α-鹅膏蕈碱的敏感性与诱导信号中的不同。这些实验有助于确定βmaj珠蛋白mRNA产生的可诱导转录单位。