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The Three Fungal Transmembrane Nuclear Pore Complex Proteins of Aspergillus nidulans Are Dispensable in the Presence of an Intact An-Nup84-120 Complex

机译:在完整的An-Nup84-120复合物存在下,构巢曲霉的三种真菌跨膜核孔复合物蛋白是可有可无的。

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摘要

In Aspergillus nidulans nuclear pore complexes (NPCs) undergo partial mitotic disassembly such that 12 NPC proteins (Nups) form a core structure anchored across the nuclear envelope (NE). To investigate how the NPC core is maintained, we affinity purified the major core An-Nup84-120 complex and identified two new fungal Nups, An-Nup37 and An-ELYS, previously thought to be vertebrate specific. During mitosis the An-Nup84-120 complex locates to the NE and spindle pole bodies but, unlike vertebrate cells, does not concentrate at kinetochores. We find that mutants lacking individual An-Nup84-120 components are sensitive to the membrane destabilizer benzyl alcohol (BA) and high temperature. Although such mutants display no defects in mitotic spindle formation, they undergo mitotic specific disassembly of the NPC core and transient aggregation of the mitotic NE, suggesting the An-Nup84-120 complex might function with membrane. Supporting this, we show cells devoid of all known fungal transmembrane Nups (An-Ndc1, An-Pom152, and An-Pom34) are viable but that An-ndc1 deletion combined with deletion of individual An-Nup84-120 components is either lethal or causes sensitivity to treatments expected to destabilize membrane. Therefore, the An-Nup84-120 complex performs roles, perhaps at the NPC membrane as proposed previously, that become essential without the An-Ndc1 transmembrane Nup.
机译:在构巢曲霉中,核孔复合体(NPC)经历部分有丝分裂拆卸,从而使12个NPC蛋白(Nups)形成锚定在核膜层(NE)上的核心结构。为了研究如何维持NPC核心,我们亲和纯化了主要的核心An-Nup84-120复合物,并鉴定了两个新的真菌小坚果An-Nup37和An-ELYS,以前认为它们是脊椎动物特有的。在有丝分裂期间,An-Nup84-120复合体位于NE和纺锤体上,但与脊椎动物细胞不同,它不集中在动植物上。我们发现缺少单个An-Nup84-120组件的突变体对膜去稳定剂苄醇(BA)和高温敏感。尽管此类突变体在有丝分裂纺锤体形成中未显示缺陷,但它们会经历NPC核的有丝分裂特异性分解和有丝分裂NE的短暂聚集,表明An-Nup84-120复合物可能与膜一起起作用。支持这一点,我们显示缺少所有已知的真菌跨膜小瘤(An-Ndc1,An-Pom152和An-Pom34)的细胞是可行的,但An-ndc1缺失与单个An-Nup84-120组分的缺失相结合是致命的或对预期会使膜不稳定的治疗敏感。因此,An-Nup84-120复合物可能发挥作用,也许是在先前提出的NPC膜上,而没有An-Ndc1跨膜Nup则变得至关重要。

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