Rasheed Zeshaan A, Saleem Ahamed, Ravee Yaniv, Pandolfi Pier Paolo, Rubin Eric H
Department of Pharmacology, Robert Wood Johnson Medical School, New Brunswick, New Jersey 08903-2681, USA.
Exp Cell Res. 2002 Jul 15;277(2):152-60. doi: 10.1006/excr.2002.5550.
We previously identified topors as a topoisomerase I-binding protein that localizes in punctate nuclear regions when expressed as a GFP fusion protein. We now demonstrate that both the GFP-topors fusion protein and endogenous topors are associated with promyelocytic leukemia (PML) nuclear bodies in exponentially growing HeLa cells. Studies using isogenic PML+/+ and PML-/- murine embryonic fibroblasts indicate that the punctate nuclear localization of topors is dependent on PML. A basic C-terminal region but not the N-terminal RING domain of topors is required for the punctate nuclear localization of this protein. Additional studies indicate that topors, but not PML, rapidly relocalizes from nuclear bodies to the nucleoplasm in cells exposed to the transcription inhibitor dichloro-1-beta-d-ribofuranolsylbenzimidazole or to the topoisomerase I-targeting drug camptothecin. These results identify topors as a new member of the group of proteins that associate dynamically with PML nuclear bodies and suggest that topors may be involved in the cellular response to camptothecin.
我们之前鉴定出topors是一种拓扑异构酶I结合蛋白,当作为绿色荧光蛋白(GFP)融合蛋白表达时,定位于点状核区域。我们现在证明,在指数生长的HeLa细胞中,GFP-topors融合蛋白和内源性topors均与早幼粒细胞白血病(PML)核体相关。使用同基因PML+/+和PML-/-小鼠胚胎成纤维细胞的研究表明,topors的点状核定位依赖于PML。该蛋白的点状核定位需要topors的碱性C末端区域而非N末端RING结构域。进一步的研究表明,在暴露于转录抑制剂二氯-1-β-D-呋喃核糖基苯并咪唑或拓扑异构酶I靶向药物喜树碱的细胞中,topors而非PML会迅速从核体重新定位到核质中。这些结果确定topors是与PML核体动态相关的蛋白质组的新成员,并表明topors可能参与细胞对喜树碱的反应。