Cvacková Zuzana, Albring Kai F, Koberna Karel, Ligasová Anna, Huber Otmar, Raska Ivan, Stanek David
Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 128 00 Prague 2, Czech Republic.
Chromosoma. 2008 Oct;117(5):487-97. doi: 10.1007/s00412-008-0170-8. Epub 2008 Jun 12.
Pontin is a multifunctional protein having roles in various cellular processes including regulation of gene expression. Here, we addressed Pontin intracellular localization using two different monoclonal antibodies directed against different Pontin epitopes. For the first time, Pontin was directly visualized in nucleoli where it co-localizes with Upstream Binding Factor and RNA polymerase I. Nucleolar localization of Pontin was confirmed by its detection in nucleolar extracts and by electron microscopy, which revealed Pontin accumulation specifically in the nucleolar fibrillar centers. Pontin localization in the nucleolus was dynamic and Pontin accumulated in large nucleolar dots mainly during S-phase. Pontin concentration in the large nucleolar dots correlated with reduced transcriptional activity of nucleoli. In addition, Pontin was found to associate with RNA polymerase I and to interact in a complex with c-Myc with rDNA sequences indicating that Pontin is involved in the c-Myc-dependent regulation of rRNA synthesis.
Pontin是一种多功能蛋白质,在包括基因表达调控在内的各种细胞过程中发挥作用。在此,我们使用两种针对不同Pontin表位的不同单克隆抗体研究了Pontin在细胞内的定位。首次在核仁中直接观察到Pontin,它与上游结合因子和RNA聚合酶I共定位。通过在核仁提取物中的检测以及电子显微镜观察证实了Pontin在核仁中的定位,电子显微镜显示Pontin特异性积聚在核仁纤维中心。Pontin在核仁中的定位是动态的,并且Pontin主要在S期积聚在大的核仁点中。大核仁点中的Pontin浓度与核仁转录活性降低相关。此外,发现Pontin与RNA聚合酶I相关,并与c-Myc在一个与rDNA序列形成的复合物中相互作用,表明Pontin参与了c-Myc依赖的rRNA合成调控。