Dundr M, Misteli T, Olson M O
Department of Biochemistry, University of Mississippi Medical Center, Jackson Mississippi 39216, USA.
J Cell Biol. 2000 Aug 7;150(3):433-46. doi: 10.1083/jcb.150.3.433.
Mammalian cell nucleoli disassemble at the onset of M-phase and reassemble during telophase. Recent studies showed that partially processed preribosomal RNA (pre-rRNA) is preserved in association with processing components in the perichromosomal regions (PRs) and in particles called nucleolus-derived foci (NDF) during mitosis. Here, the dynamics of nucleolar reassembly were examined for the first time in living cells expressing fusions of the processing-related proteins fibrillarin, nucleolin, or B23 with green fluorescent protein (GFP). During telophase the NDF disappeared with a concomitant appearance of material in the reforming nuclei. Prenucleolar bodies (PNBs) appeared in nuclei in early telophase and gradually disappeared as nucleoli formed, strongly suggesting the transfer of PNB components to newly forming nucleoli. Fluorescence recovery after photobleaching (FRAP) showed that fibrillarin-GFP reassociates with the NDF and PNBs at rapid and similar rates. The reentry of processing complexes into telophase nuclei is suggested by the presence of pre-rRNA sequences in PNBs. Entry of specific proteins into the nucleolus approximately correlated with the timing of processing events. The mitotically preserved processing complexes may be essential for regulating the distribution of components to reassembling daughter cell nucleoli.
哺乳动物细胞的核仁在M期开始时解体,并在末期重新组装。最近的研究表明,在有丝分裂期间,部分加工的前核糖体RNA(pre-rRNA)与加工成分一起保留在染色体周围区域(PRs)和称为核仁衍生灶(NDF)的颗粒中。在此,首次在表达加工相关蛋白纤维蛋白原、核仁蛋白或B23与绿色荧光蛋白(GFP)融合体的活细胞中研究了核仁重新组装的动力学。在末期,NDF消失,同时在重新形成的细胞核中出现物质。核仁前体(PNBs)在末期早期出现在细胞核中,并随着核仁的形成逐渐消失,这强烈表明PNB成分转移到了新形成的核仁中。光漂白后荧光恢复(FRAP)显示,纤维蛋白原-GFP以快速且相似的速率与NDF和PNBs重新结合。PNBs中存在pre-rRNA序列表明加工复合物重新进入末期细胞核。特定蛋白质进入核仁的时间大致与加工事件的时间相关。有丝分裂期间保留的加工复合物可能对于调节成分向重新组装的子细胞核仁的分布至关重要。