Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy.
Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy.
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DOI：
10.1186/s13024-022-00585-1
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发表时间：
2022-12-08
影响因子：
15.1
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中科院分区：
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TAR DNA结合蛋白-43(TDP-43)的细胞质错误定位和聚集是肌萎缩侧索硬化症和额颞叶痴呆(ALS/FTD)疾病谱的标志，导致核功能丧失和细胞质毒性功能获得表型。虽然TDP-43蛋白病与核质转运缺陷有关，但这一过程仍然知之甚少。在这里，我们研究了核粘附素-β1(KPNB1)和其他核输入受体在调节TDP-43病理中的作用。我们在细胞系、原代神经元和器官型小鼠脑片培养中使用免疫染色、免疫沉淀、生化和毒性分析，以确定KPNB1对病理性TDP-43结构的溶解性、定位和毒性的影响。尸检患者的脑和脊髓组织染色，以评估KPNB1与TDP-43包涵体的共定位。用比浊法研究了重组TDP-43纤维的体外溶解和防止聚集的作用。采用果蝇TDP-43蛋白病模型，观察KPNB1对其体内神经退行性变表型的影响。我们发现，核输入受体蛋白家族的几个成员可以减少病理性TDP-43聚集体的形成。以KPNB1为模型，我们发现它的活性依赖于TDP-43的Pron样C-末端区域，该区域介导了与苯丙氨酸和富含甘氨酸的核孔蛋白(FG-nup)如Nup62的共聚集。KPNB1被招募到这些共聚集体中，在那里它作为分子伴侣逆转Nup62和TDP-43的异常相变。这些发现得到了以下发现的支持：在ALS/FTD死后的CNS组织中，Nup62和KPNB1也被隔离在病理性的TDP-43聚集体中，并且KPNB1的苍蝇同源基因在TDP-43蛋白病的果蝇模型中被鉴定为一种强大的保护性修饰物。我们的结果表明，KPNB1可以恢复TDP-43的溶解性和核定位，并减少ALS/FTD细胞和动物模型中的神经变性，从而挽救TDP-43病理的所有特征。我们的发现表明了一种新的NLS非依赖机制，与其在溶解核孔中FG-NUP形成的扩散屏障方面的典型作用类似，KPNB1被招募到TDP-43蛋白病变中存在的TDP-43/FG-Nup共聚集体中，并在治疗上逆转其有害的相变和错误定位，减轻神经变性。网上版载有补充材料，可在10.1186/s13024-022-00585-1查阅。
Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. While TDP-43 proteinopathy has been associated with defects in nucleocytoplasmic transport, this process is still poorly understood. Here we study the role of karyopherin-β1 (KPNB1) and other nuclear import receptors in regulating TDP-43 pathology. We used immunostaining, immunoprecipitation, biochemical and toxicity assays in cell lines, primary neuron and organotypic mouse brain slice cultures, to determine the impact of KPNB1 on the solubility, localization, and toxicity of pathological TDP-43 constructs. Postmortem patient brain and spinal cord tissue was stained to assess KPNB1 colocalization with TDP-43 inclusions. Turbidity assays were employed to study the dissolution and prevention of aggregation of recombinant TDP-43 fibrils in vitro. Fly models of TDP-43 proteinopathy were used to determine the effect of KPNB1 on their neurodegenerative phenotype in vivo. We discovered that several members of the nuclear import receptor protein family can reduce the formation of pathological TDP-43 aggregates. Using KPNB1 as a model, we found that its activity depends on the prion-like C-terminal region of TDP-43, which mediates the co-aggregation with phenylalanine and glycine-rich nucleoporins (FG-Nups) such as Nup62. KPNB1 is recruited into these co-aggregates where it acts as a molecular chaperone that reverses aberrant phase transition of Nup62 and TDP-43. These findings are supported by the discovery that Nup62 and KPNB1 are also sequestered into pathological TDP-43 aggregates in ALS/FTD postmortem CNS tissue, and by the identification of the fly ortholog of KPNB1 as a strong protective modifier in Drosophila models of TDP-43 proteinopathy. Our results show that KPNB1 can rescue all hallmarks of TDP-43 pathology, by restoring its solubility and nuclear localization, and reducing neurodegeneration in cellular and animal models of ALS/FTD. Our findings suggest a novel NLS-independent mechanism where, analogous to its canonical role in dissolving the diffusion barrier formed by FG-Nups in the nuclear pore, KPNB1 is recruited into TDP-43/FG-Nup co-aggregates present in TDP-43 proteinopathies and therapeutically reverses their deleterious phase transition and mislocalization, mitigating neurodegeneration. The online version contains supplementary material available at 10.1186/s13024-022-00585-1.
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影响因子：
25
作者：
Chou CC;Zhang Y;Umoh ME;Vaughan SW;Lorenzini I;Liu F;Sayegh M;Donlin-Asp PG;Chen YH;Duong DM;Seyfried NT;Powers MA;Kukar T;Hales CM;Gearing M;Cairns NJ;Boylan KB;Dickson DW;Rademakers R;Zhang YJ;Petrucelli L;Sattler R;Zarnescu DC;Glass JD;Rossoll W
通讯作者：
Rossoll W
影响因子：
5
作者：
Carey JL;Guo L
通讯作者：
Guo L
影响因子：
64.8
作者：
Arseni D;Hasegawa M;Murzin AG;Kametani F;Arai M;Yoshida M;Ryskeldi-Falcon B
通讯作者：
Ryskeldi-Falcon B
影响因子：
17.1
作者：
Cook CN;Wu Y;Odeh HM;Gendron TF;Jansen-West K;Del Rosso G;Yue M;Jiang P;Gomes E;Tong J;Daughrity LM;Avendano NM;Castanedes-Casey M;Shao W;Oskarsson B;Tomassy GS;McCampbell A;Rigo F;Dickson DW;Shorter J;Zhang YJ;Petrucelli L
通讯作者：
Petrucelli L
影响因子：
15.1
作者：
Chew, Jeannie;Cook, Casey;Petrucelli, Leonard
通讯作者：
Petrucelli, Leonard