Silencing of OSBP-related protein 8 (ORP8) modifies the macrophage transcriptome, nucleoporin p62 distribution, and migration capacity
Silencing of OSBP-related protein 8 (ORP8) modifies the macrophage transcriptome, nucleoporin p62 distribution, and migration capacity
复制标题
OSBP 相关蛋白 8 (ORP8) 的沉默会改变巨噬细胞转录组、核孔蛋白 p62 分布和迁移能力
DOI：
10.1016/j.yexcr.2012.05.026
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发表时间：
2012-09-10
影响因子：
3.7
通讯作者：
Olkkonen, Vesa M.
中科院分区：
文献类型：
--
作者：
Beaslas, Olivier;Vihervaara, Terhi;Olkkonen, Vesa M.
ORP8 是一种锚定在内质网和核膜上的氧甾醇/胆固醇结合蛋白，在巨噬细胞中大量表达。我们使用 shRNA 慢病毒创建并表征了 ORP8 稳定沉默的小鼠 RAW264.7 巨噬细胞。微阵列转录组和基因本体通路分析揭示了几种核通路以及与中心体和微管组织相关的通路的显着改变。 ORP8 敲低导致 ORP8 的相互作用伴侣核孔蛋白 NUP62 的表达增加并改变亚细胞分布，具有核内定位方面以及与细胞质囊泡结构和细胞片状边缘的关联。此外，与表达非靶向shRNA的对照组相比，ORP8沉默的细胞表现出增强的迁移和更明显的微管细胞骨架。 ORP8 与 Exo70 竞争与 NUP62 的相互作用，并且 NUP62 敲除消除了 ORP8 沉默细胞的迁移增强，表明内源性 ORP8 通过与 NUP62 结合来抑制迁移。总之，本研究揭示了巨噬细胞中ORP8功能的新的、意想不到的方面，不直接涉及脂质代谢，而是与核功能、微管组织和迁移能力相关。 (C) 2012 Elsevier Inc. 保留所有权利。
ORP8 is an oxysterol/cholesterol binding protein anchored to the endoplasmic reticulum and the nuclear envelope, and is abundantly expressed in the macrophage. We created and characterized mouse RAW264.7 macrophages with ORP8 stably silenced using shRNA lentiviruses. A microarray transcriptome and gene ontology pathway analysis revealed significant alterations in several nuclear pathways and ones associated with centrosome and microtubule organization. ORP8 knockdown resulted in increased expression and altered subcellular distribution of an interaction partner of ORP8, nucleoporin NUP62, with an intranuclear localization aspect and association with cytoplasmic vesicular structures and lamellipodial edges of the cells. Moreover, ORP8 silenced cells displayed enhanced migration, and a more pronounced microtubule cytoskeleton than controls expressing a non-targeting shRNA. ORP8 was shown to compete with Exo70 for interaction with NUP62, and NUP62 knockdown abolished the migration enhancement of ORP8-silenced cells, suggesting that the endogenous ORP8 suppresses migration via binding to NUP62. As a conclusion, the present study reveals new, unexpected aspects of ORP8 function in macrophages not directly involving lipid metabolism, but rather associated with nuclear functions, microtubule organization, and migration capacity. (C) 2012 Elsevier Inc. All rights reserved.