Energy- and temperature-dependent transport of integral proteins to the inner nuclear membrane via the nuclear pore.

Energy- and temperature-dependent transport of integral proteins to the inner nuclear membrane via the nuclear pore.
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DOI:
10.1083/jcb.200409149
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发表时间:
2004-12-20
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Gerace L
Gerace L
中科院分区:
其他
文献类型:
--
作者:
Ohba T;Schirmer EC;Nishimoto T;Gerace L

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内核膜(INM)的常驻整合蛋白在外周内质网(ER)上合成为膜整合蛋白,并使用未知的运输机制在整个间期运输到INM。为了研究这种转运,我们开发了一种活细胞测定法,该测定法通过雷帕霉素介导的在核纤层处的捕获来测量跨膜报告分子从ER到INM的移动。具有小(<30 kD)胞质和内腔结构域的报告基因构建体在INM处快速积累。然而,增加47 kD的任何域的大小强烈抑制运动。降低温度和ATP耗竭也抑制运动,这是膜融合机制的特征,但药物抑制囊泡运输没有影响。由于报告的积累在INM的抑制抗体的核孔膜蛋白gp210,我们的研究结果支持一个模型,其中运输的整合蛋白质的INM涉及在脂双层的核孔膜周围的横向扩散,加上积极重组的核孔复合物。
Resident integral proteins of the inner nuclear membrane (INM) are synthesized as membrane-integrated proteins on the peripheral endoplasmic reticulum (ER) and are transported to the INM throughout interphase using an unknown trafficking mechanism. To study this transport, we developed a live cell assay that measures the movement of transmembrane reporters from the ER to the INM by rapamycin-mediated trapping at the nuclear lamina. Reporter constructs with small (<30 kD) cytosolic and lumenal domains rapidly accumulated at the INM. However, increasing the size of either domain by 47 kD strongly inhibited movement. Reduced temperature and ATP depletion also inhibited movement, which is characteristic of membrane fusion mechanisms, but pharmacological inhibition of vesicular trafficking had no effect. Because reporter accumulation at the INM was inhibited by antibodies to the nuclear pore membrane protein gp210, our results support a model wherein transport of integral proteins to the INM involves lateral diffusion in the lipid bilayer around the nuclear pore membrane, coupled with active restructuring of the nuclear pore complex.
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