Chen D, Huang S
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
J Cell Biol. 2001 Apr 2;153(1):169-76. doi: 10.1083/jcb.153.1.169.
We examined the mobilities of nucleolar components that act at various steps of the ribosome biogenesis pathway. Fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) analyses demonstrate that factors involved in rRNA transcription (upstream-binding factor [UBF]), processing (nucleolin, fibrillarin, and RNase MRP subunits, Rpp29), and ribosome assembly (B23) exchange rapidly between the nucleoplasm and nucleolus. In contrast, the mobilities of ribosomal subunit proteins (S5, L9) are much slower. Selective inhibition of RNA polymerase I transcription does not prevent the exchanges but influences the rates of exchange differentially for different nucleolar components. These findings suggest that the rapid exchange of nucleolar components between the nucleolus and nucleoplasm may represent a new level of regulation for rRNA synthesis. The different dynamic properties of proteins involved in different steps of ribosome biogenesis imply that the nucleolar association of these proteins is due to their specific functional roles rather than simply their specific nucleolar-targeting events.
我们研究了在核糖体生物合成途径不同步骤发挥作用的核仁组分的流动性。光漂白后荧光恢复(FRAP)和光漂白荧光损失(FLIP)分析表明,参与rRNA转录(上游结合因子[UBF])、加工(核仁素、纤维蛋白原和核糖核酸酶MRP亚基Rpp29)以及核糖体组装(B23)的因子在核质和核仁之间快速交换。相比之下,核糖体亚基蛋白(S5、L9)的流动性要慢得多。RNA聚合酶I转录的选择性抑制并不阻止交换,但对不同核仁组分的交换速率有不同影响。这些发现表明,核仁组分在核仁和核质之间的快速交换可能代表了rRNA合成调控的一个新水平。参与核糖体生物合成不同步骤的蛋白质具有不同的动态特性,这意味着这些蛋白质与核仁的结合是由于它们特定的功能作用,而不仅仅是它们特定的核仁靶向事件。