Zatsepina O V, Voit R, Grummt I, Spring H, Semenov M V, Trendelenburg M F
A. N. Belozersky Institute of Physical and Chemical Biology, Moscow State University, Russia.
Chromosoma. 1993 Nov;102(9):599-611. doi: 10.1007/BF00352307.
We have characterized an anti-NOR (nucleolar organizer region) serum (P419) from a patient with rheumatoid arthritis and show that it contains antibodies directed against the RNA polymerase I-specific transcription initiation factor UBF. This serum reacts with UBF from a variety of vertebrate cells as revealed both by immunoblotting and by indirect immunofluorescence. We have used the P419 serum to study the intracellular localization of this transcription factor at the light and electron microscopic level. In interphase cells, UBF exhibits a pronounced punctate pattern and is found to be associated with necklace-like structures, which appear to reflect the transcriptionally active state of the nucleolus. Inhibition of rRNA synthetic activity caused either by nutritional starvation or by actinomycin D treatment resulted in a marked decrease in the number and in a significant increase in the size of UBF-positive granules. Under all experimental conditions applied, UBF was exclusively found within the nucleolus and was not released into the nucleoplasm or cytoplasm. During mitosis, UBF was found to be concentrated at the chromosomal NOR indicating that a significant quantity, if not all, of this factor remains bound to the ribosomal transcription units. From this we conclude that UBF is associated both with transcriptionally active and inactive rRNA genes and, therefore, changes in the intracellular localization of UBF are very likely not involved in rDNA transcription regulation.
我们对一名类风湿性关节炎患者的抗核仁组织区(NOR)血清(P419)进行了特性分析,结果表明该血清含有针对RNA聚合酶I特异性转录起始因子UBF的抗体。免疫印迹和间接免疫荧光分析均显示,该血清能与多种脊椎动物细胞中的UBF发生反应。我们利用P419血清在光学和电子显微镜水平上研究了这种转录因子的细胞内定位。在间期细胞中,UBF呈现出明显的点状模式,并且发现与项链样结构相关,这些结构似乎反映了核仁的转录活性状态。营养饥饿或放线菌素D处理导致的rRNA合成活性抑制,使得UBF阳性颗粒的数量显著减少,而颗粒大小则显著增加。在所有应用的实验条件下,UBF仅存在于核仁内,并未释放到核质或细胞质中。在有丝分裂期间,发现UBF集中在染色体NOR处,这表明该因子即便不是全部,也有相当数量仍然与核糖体转录单元结合。由此我们得出结论,UBF与转录活性和非活性的rRNA基因均有关联,因此,UBF细胞内定位的变化很可能与rDNA转录调控无关。