ER membrane-bending proteins are necessary for de novo nuclear pore formation.

ER membrane-bending proteins are necessary for de novo nuclear pore formation.
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DOI:
10.1083/jcb.200806174
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发表时间:
2009-03-09
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Wente SR
Wente SR
中科院分区:
其他
文献类型:
--
作者:
Dawson TR;Lazarus MD;Hetzer MW;Wente SR

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核质转运仅通过核孔复合物(NPC)发生,核孔复合物嵌入由内外核膜融合形成的孔中。新生孔和NPC生物发生的机制仍不清楚。Reticulons(RTNs)和Yop 1/DP 1是形成和维持管状内质网(ER)和有丝分裂后核膜所需的保守膜蛋白家族。在这项研究中,我们报告说,RTN和Yop 1/DP 1家族的成员是核孔形成所需的。对酿酒酵母prp 20-G282 S和nup 133 Δ NPC组装突变体的分析揭示了Rtn 1-绿色荧光蛋白(GFP)和Yop 1-GFP ER分布和共定位于NPC簇的扰动。RTN 1和YOP 1的组合缺失导致NPC聚集、核输入缺陷和合成致死性,另外缺乏Pom 34、Pom 152和Nup 84亚复合体成员。我们使用非洲爪蟾体外试验测试了NPC生物发生中的直接作用,发现抗Rtn 4a抗体特异性抑制从头核孔形成。我们假设这些ER膜弯曲蛋白介导早期NPC组装步骤。
Nucleocytoplasmic transport occurs exclusively through nuclear pore complexes (NPCs) embedded in pores formed by inner and outer nuclear membrane fusion. The mechanism for de novo pore and NPC biogenesis remains unclear. Reticulons (RTNs) and Yop1/DP1 are conserved membrane protein families required to form and maintain the tubular endoplasmic reticulum (ER) and the postmitotic nuclear envelope. In this study, we report that members of the RTN and Yop1/DP1 families are required for nuclear pore formation. Analysis of Saccharomyces cerevisiae prp20-G282S and nup133Δ NPC assembly mutants revealed perturbations in Rtn1–green fluorescent protein (GFP) and Yop1-GFP ER distribution and colocalization to NPC clusters. Combined deletion of RTN1 and YOP1 resulted in NPC clustering, nuclear import defects, and synthetic lethality with the additional absence of Pom34, Pom152, and Nup84 subcomplex members. We tested for a direct role in NPC biogenesis using Xenopus laevis in vitro assays and found that anti-Rtn4a antibodies specifically inhibited de novo nuclear pore formation. We hypothesize that these ER membrane–bending proteins mediate early NPC assembly steps.
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