Miesfeld R, Sollner-Webb B, Croce C, Arnheim N
Mol Cell Biol. 1984 Jul;4(7):1306-12. doi: 10.1128/mcb.4.7.1306-1312.1984.
The basis for nucleolar dominance in mouse-human cell hybrids which contained all of the mouse chromosomes but an incomplete set of human chromosomes (M greater than H) was examined at the molecular level. S1 mapping data showed that these cells had the expected levels of steady-state rRNA transcribed from mouse ribosomal gene (rDNA) transcription units but undetectable levels of rRNA derived from the human rDNA transcription templates that are also present. RNA polymerase I-dependent, cell-free transcription extracts were made from three hybrid lines and were found to be capable of transcribing cloned rDNA templates of mouse but not human origin. Partially purified human factors required for rDNA transcription in vitro were added to the M greater than H extracts. One fraction with almost no RNA polymerase I activity conferred on these hybrid cell extracts the ability to transcribe a human rDNA template. These rescue experiments suggested that this required human-specific rDNA transcription factor(s) was effectively absent from the lines we examined and could account for nucleolar dominance in M greater than H hybrid cells.
在含有所有小鼠染色体但人类染色体组不完整(小鼠>人类)的小鼠 - 人类细胞杂交体中,从分子水平研究了核仁显性的基础。S1图谱数据表明,这些细胞中从小鼠核糖体基因(rDNA)转录单元转录的稳态rRNA水平符合预期,但来自同样存在的人类rDNA转录模板的rRNA水平却检测不到。从三个杂交系制备了依赖RNA聚合酶I的无细胞转录提取物,发现它们能够转录小鼠而非人类来源的克隆rDNA模板。将体外rDNA转录所需的部分纯化的人类因子添加到小鼠>人类的提取物中。一种几乎没有RNA聚合酶I活性的组分赋予这些杂交细胞提取物转录人类rDNA模板的能力。这些拯救实验表明,我们检测的细胞系中有效缺乏这种所需的人类特异性rDNA转录因子,这可以解释小鼠>人类杂交细胞中的核仁显性现象。