Schul W, van Driel R, de Jong L
E.C. Slater Instituut, University of Amsterdam, BioCentrum Amsterdam, 1018 TV Amsterdam, The Netherlands.
Mol Biol Cell. 1998 May;9(5):1025-36. doi: 10.1091/mbc.9.5.1025.
A significant percentage of the gene clusters that contain the human genes for U1 small nuclear RNA (snRNA) or for U2 snRNA have been found associated with small nuclear domains, known as coiled bodies. We show here, by immunofluorescent labeling of human cells, that coiled bodies are enriched in factors required for the transcription of these snRNA genes. The 45-kDa gamma-subunit of the transcription factor, proximal element sequence-binding transcription factor (PTF), which is specific for the snRNA genes, was found in high concentrations in coiled bodies, along with the general transcription factor TATA-box binding protein and a subset of RNA polymerase II. We show that the transcription factors and RNA polymerase II are concentrated in irregularly shaped domains that not only overlap with coiled bodies but also extend to their immediate surroundings. Fluorescent in situ hybridization showed that these domains can overlap with U2 snRNA genes adjacent to coiled bodies. In addition, we found the domains to contain newly synthesized RNA, visualized by 5-bromo-uridine triphosphate labeling. Our data suggest that coiled bodies are involved in the expression of snRNA genes, which leads us to propose the model that coiled bodies are associated with snRNA genes to facilitate and regulate their transcription. These findings point to a general principle of higher order organization of gene expression in the nucleus.
已发现相当比例的包含人类U1小核RNA(snRNA)基因或U2 snRNA基因的基因簇与称为卷曲小体的小核结构域相关。我们通过对人类细胞进行免疫荧光标记表明,卷曲小体富含这些snRNA基因转录所需的因子。转录因子近端元件序列结合转录因子(PTF)的45 kDaγ亚基对snRNA基因具有特异性,它与通用转录因子TATA盒结合蛋白以及RNA聚合酶II的一个亚群一起在卷曲小体中高浓度存在。我们表明,转录因子和RNA聚合酶II集中在不规则形状的结构域中,这些结构域不仅与卷曲小体重叠,还延伸到其紧邻区域。荧光原位杂交显示,这些结构域可与卷曲小体附近的U2 snRNA基因重叠。此外,我们发现这些结构域含有通过5-溴尿苷三磷酸标记可视化的新合成RNA。我们的数据表明卷曲小体参与snRNA基因的表达,这使我们提出卷曲小体与snRNA基因相关联以促进和调节其转录的模型。这些发现指出了细胞核中基因表达高阶组织的一般原则。