Endogenous RGS14 is a cytoplasmic-nuclear shuttling protein that localizes to juxtanuclear membranes and chromatin-rich regions of the nucleus.
Endogenous RGS14 is a cytoplasmic-nuclear shuttling protein that localizes to juxtanuclear membranes and chromatin-rich regions of the nucleus.
复制标题
DOI：
10.1371/journal.pone.0184497
复制
发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hepler JR
中科院分区：
文献类型：
--
作者：
Branch MR;Hepler JR
G蛋白信号调节因子14（Regulator of G protein signaling 14，RGS 14）是一种多功能支架蛋白，它整合了G蛋白和H-Ras/MAP激酶信号通路，调节海马学习记忆过程中的突触可塑性。然而，迄今为止，很少有人知道的亚细胞分布和内源性RGS 14在神经元细胞系中的作用。大多数关于RGS 14细胞行为的已知是基于在非天然宿主细胞中异位过表达的标记的重组RGS 14的研究。在这里，我们报告的第一次全面评估的亚细胞分布和动态定位的内源性RGS 14在大鼠B35神经母细胞瘤细胞。使用共聚焦成像和3D结构照明显微镜，我们发现内源性RGS 14定位于亚细胞区室，以前没有在重组RGS 14的研究中认识到。RGS 14的定位观察到最显着的包围细胞核，在核膜两侧的核孔复合物（NPC）和核内膜通道内，并在细胞周期依赖性的方式在两个染色质贫乏和染色质丰富的区域的核。此外，一个子集的核RGS 14定位邻近活性RNA聚合酶II。在静息细胞中，质膜不存在内源性RGS 14;然而，在使用AlF 4 '组成性激活内源性RGS 14 G蛋白结合伴侣后，该蛋白可以从ER/高尔基体衍生的内体中的非核膜运输到质膜。最后，我们的研究结果表明，内源性RGS 14的行为作为一个细胞质-核穿梭蛋白证实了什么以前已经显示的重组RGS 14。总之，这些发现突出了先前未认识到的RGS 14可能的细胞作用，这些作用与常规GPCR-G蛋白信号传导的调节不同，特别是RGS 14在细胞核中的未定义作用。
Regulator of G protein signaling 14 (RGS14) is a multifunctional scaffolding protein that integrates G protein and H-Ras/MAPkinase signaling pathways to regulate synaptic plasticity important for hippocampal learning and memory. However, to date, little is known about the subcellular distribution and roles of endogenous RGS14 in a neuronal cell line. Most of what is known about RGS14 cellular behavior is based on studies of tagged, recombinant RGS14 ectopically overexpressed in unnatural host cells. Here, we report for the first time a comprehensive assessment of the subcellular distribution and dynamic localization of endogenous RGS14 in rat B35 neuroblastoma cells. Using confocal imaging and 3D-structured illumination microscopy, we find that endogenous RGS14 localizes to subcellular compartments not previously recognized in studies of recombinant RGS14. RGS14 localization was observed most notably at juxtanuclear membranes encircling the nucleus, at nuclear pore complexes (NPC) on both sides of the nuclear envelope and within intranuclear membrane channels, and within both chromatin-poor and chromatin-rich regions of the nucleus in a cell cycle-dependent manner. In addition, a subset of nuclear RGS14 localized adjacent to active RNA polymerase II. Endogenous RGS14 was absent from the plasma membrane in resting cells; however, the protein could be trafficked to the plasma membrane from juxtanuclear membranes in endosomes derived from ER/Golgi, following constitutive activation of endogenous RGS14 G protein binding partners using AlF4¯. Finally, our findings show that endogenous RGS14 behaves as a cytoplasmic-nuclear shuttling protein confirming what has been shown previously for recombinant RGS14. Taken together, the findings highlight possible cellular roles for RGS14 not previously recognized that are distinct from the regulation of conventional GPCR-G protein signaling, in particular undefined roles for RGS14 in the nucleus.
登录
查看更多内容
DOI：
10.1083/jcb.136.3.531
发表时间：
1997-02-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fricker M;Hollinshead M;White N;Vaux D
通讯作者：
Vaux D
影响因子：
16.2
作者：
Frank, Christopher L.;Tsai, Li-Huei
通讯作者：
Tsai, Li-Huei
影响因子：
7.8
作者：
Cho, Hyeseon;Kehrl, John H.
通讯作者：
Kehrl, John H.
影响因子：
4.8
作者：
Brown, Nicole E.;Goswami, Devrishi;Hepler, John R.
通讯作者：
Hepler, John R.
影响因子：
3.6
作者：
Cho, H;Kozasa, T;Kehrl, JH
通讯作者：
Kehrl, JH