Smirnova O Iu, Dudnik O A, Zatsepina O V
A. N. Belozersky Institute of Physical and Chemical Biology, Moscow University.
Tsitologiia. 2002;44(1):5-13.
It is well known that fibrillar centers (FC) constitute an essential structural component of the active nucleolus in mammalian cells, yet their role in regulation of ribosomal gene transcription still remains an open question. Here, we studied the activity of endogenous RNA polymerase I upon partial and complete unraveling of nucleoli and FCs. The pattern of BrUTP incorporation in nuclei of hypotonically-treated cells was shown to be essentially the same as in the control untreated cells. Moreover, the sites of BrUTP incorporation, which revealed the active PNA polymerase I, were completely coincident with UBF-binding sites. These observations allow to conclude that structural integrity of FCs is not a prerequisite for maintenance of the active RNA polymerase I transcriptional complex. When the action of hypotonic shock was ceased and the cells were transferred to a complete cultural medium, the swollen nucleoli recovered to the control state. Therefore it is possible to conclude that none of the main morphological nucleolar counterparts, such as FCs, dense fibrillar component or the pars granulosa, is responsible for the maintenance of the nucleolar structural and functional integrity. A suggestion is made that this role may be played by the nucleolar matrix associated with the RNA polymerase I transcriptional complex.
众所周知,纤维中心(FC)是哺乳动物细胞中活性核仁的重要结构组成部分,但其在核糖体基因转录调控中的作用仍是一个悬而未决的问题。在此,我们研究了核仁和纤维中心部分或完全解聚时内源性RNA聚合酶I的活性。结果显示,低渗处理细胞的细胞核中溴尿嘧啶三磷酸(BrUTP)掺入模式与未处理的对照细胞基本相同。此外,揭示活性RNA聚合酶I的BrUTP掺入位点与上游结合因子(UBF)结合位点完全重合。这些观察结果表明,纤维中心的结构完整性并非维持活性RNA聚合酶I转录复合体的必要条件。当低渗休克作用停止且细胞转移至完全培养基中时,肿胀的核仁恢复至对照状态。因此,可以得出结论,核仁的主要形态对应物,如纤维中心、致密纤维组分或颗粒区,均不负责维持核仁的结构和功能完整性。有人提出,这一作用可能由与RNA聚合酶I转录复合体相关的核仁基质发挥。