Nucleolar localization of DGCR8 and identification of eleven DGCR8-associated proteins
Nucleolar localization of DGCR8 and identification of eleven DGCR8-associated proteins
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DOI：
10.1016/j.yexcr.2007.07.020
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发表时间：
2007-12-10
影响因子：
3.7
通讯作者：
Shimizu, Nobuyoshi
中科院分区：
文献类型：
--
作者：
Shiohama, Aiko;Sasaki, Takashi;Shimizu, Nobuyoshi
通过免疫沉淀和质谱分析，我们鉴定了11个与Dgcr8相关的蛋白质。这些蛋白质包括核仁素、ILF3等，其中大部分似乎参与了RNA的加工或运输。免疫共沉淀法至少检测到DROSHA/DgCr8、DgCr8/核仁素、DgCr8/ILF3和ILF3/XPO5四种蛋白质复合体。Dgcr8与核仁络合物的形成依赖于RNA。免疫荧光显微镜和免疫电子显微镜的亚细胞定位分析表明，Dgcr8位于核仁和与核质内的剪接斑点相邻的小灶处。此外，抑制RNA转录改变了Dgcr8在核仁上的定位。因此，我们的研究为各种蛋白质复合体参与表面复杂的先天RNA干扰机制的分子机制提供了额外的新证据。(C)2007 Elsevier Inc.保留所有权利
We identified 11 proteins that are associated with DGCR8 by immunoprecipitation assay and mass spectrometry. These proteins included Nucleolin, ILF3 and others, most of which appeared to be involved in the RNA processing or RNA transportation. We detected at least four kinds of protein complex, such as DROSHA/DGCR8, DGCR8/Nucleolin, DGCR8/ILF3 and ILF3/XPO5, by co-immunoprecipitation. The complex formation of DGCR8 with Nucleolin was dependent on RNA. Subcellular localization analysis by the immuno fluorescent microscopy and immunoelectron microscopy indicated that DGCR8 locates at the nucleolus and small foci adjacent to splicing speckles in the nucleoplasm. Furthermore, the localization of DGCR8 at the nucleolus was changed by the inhibition of RNA transcription. Thus, our studies provided additional new evidence for the involvement of various protein complexes in the molecular mechanisms of apparently complex innate RNA interference machinery. (c) 2007 Elsevier Inc. All rights reserved