Ciejek E M, Tsai M J, O'Malley B W
Nature. 1983;306(5943):607-9. doi: 10.1038/306607a0.
In the chicken oviduct, it has been well documented that steroid hormones stimulate the transcription of specific genes such as the ovalbumin gene. In addition to the presence of specific hormone receptors in the tissue, gene expression seems to require that target genes exist in large DNase I sensitive chromosomal domains. This structure appears necessary but not sufficient for transcriptional activation. In search of still other levels of control, we have investigated the interactions of genes with the nuclear matrix, a structure which has been implicated in DNA synthesis, transcription and RNA processing. Here we have isolated nuclear matrix and used a nondegradative method to fractionate nuclear DNA based on its preferential association with the matrix. The preparation was digested with a restriction enzyme and both matrix-bound and released DNAs were recovered. We found that only actively expressed genes were associated with the matrix. Furthermore, within a 100-kilobase (kb) DNase I sensitive chromosomal domain, only the transcribed regions were associated with the matrix. This association was shown to be reversible when hormone was withdrawn. Our results suggest that the nuclear matrix is the site of nuclear transcription and may represent another potential level of control for regulation of gene expression in the eukaryotic cell.
在鸡的输卵管中,类固醇激素刺激特定基因(如卵清蛋白基因)的转录,这一点已有充分的文献记载。除了组织中存在特定的激素受体外,基因表达似乎还要求靶基因存在于对脱氧核糖核酸酶I(DNase I)敏感的大染色体结构域中。这种结构对于转录激活似乎是必要的,但并不充分。为了寻找其他的调控水平,我们研究了基因与核基质的相互作用,核基质是一种与DNA合成、转录和RNA加工有关的结构。在这里,我们分离出了核基质,并使用一种非降解方法,根据核DNA与基质的优先结合对其进行分级分离。用一种限制性内切酶消化该制剂,回收与基质结合的DNA和释放的DNA。我们发现只有活跃表达的基因与基质相关。此外,在一个100千碱基(kb)对DNase I敏感的染色体结构域内,只有转录区域与基质相关。当去除激素时,这种结合被证明是可逆的。我们的结果表明,核基质是核转录的场所,可能代表了真核细胞中基因表达调控的另一个潜在调控水平。