Samal B, Ballal N R, Busch H
Cell Biol Int Rep. 1980 Feb;4(2):175-84. doi: 10.1016/0309-1651(80)90072-7.
Novikoff hepatoma cells, grown in monolayer cultures, when permeabilized by treatment with lysolecithin, incorporated 3H-UTP at a high rate for 2 hours at 25 degrees. The incorporation was inhibited by initiation inhibitors such as rifampicin AF/013, heparin and aurintricarboxylic acid. About 75% of RNA polymerase II, and 45% of RNA polymerase I activities were inhibited by rifampicin AF/013. In contrast, transcription in isolated nuclei was not inhibited either by rifampicin AF/013 or heparin. The permeabilized cells apparently retain the mechanisms for reinitiation in vitro and may be a useful model for studies on the effects of proteins on gene transcription.
在单层培养中生长的诺维科夫肝癌细胞,经溶血卵磷脂处理使其通透化后,在25摄氏度下能以较高速率掺入3H-UTP达2小时。掺入过程受到起始抑制剂如利福平AF/013、肝素和金精三羧酸的抑制。利福平AF/013可抑制约75%的RNA聚合酶II以及45%的RNA聚合酶I活性。相比之下,利福平AF/013或肝素均不抑制分离细胞核中的转录。通透化细胞显然保留了体外重新起始的机制,可能是研究蛋白质对基因转录影响的有用模型。