Interactome screening of C9orf72 dipeptide repeats reveals VCP sequestration and functional impairment by polyGA.
Interactome screening of C9orf72 dipeptide repeats reveals VCP sequestration and functional impairment by polyGA.
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DOI：
10.1093/brain/awab300
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发表时间：
2022-04-18
期刊：
影响因子：
14.5
通讯作者：
Rogelj, Boris
中科院分区：
文献类型：
--
作者：
Bozic, Janja;Motaln, Helena;Janez, Anja Pucer;Markic, Lara;Tripathi, Priyanka;Yamoah, Alfred;Aronica, Eleonora;Lee, Youn-Bok;Heilig, Raphael;Fischer, Roman;Thompson, Andrew J.;Goswami, Anand;Rogelj, Boris
C9orf72基因的重复扩增是肌萎缩性侧索硬化症和额颞叶变性的常见原因，这是两种毁灭性的神经退行性疾病。被提出的GGGGCC重复扩增的机制之一是它们被翻译成非规范的二肽重复序列，然后这些重复序列可以作为聚集体积累并导致这些病理。有五种不同的二肽重复蛋白（polyGA, polyGR, polyypr， polyypa和polyGP），其中一些已知具有神经毒性。在本研究中，我们使用BioID2接近标记来鉴定所有五种双肽重复蛋白的相互作用组，每个蛋白由125个重复组成。我们确定了polyGR的113个相互作用伙伴，polyGA的90个，polyypr的106个，polyypa的25个和polyGP的27个。对蛋白质组学数据进行基因本体富集分析发现，这些靶相互作用伙伴参与多种功能，包括蛋白质翻译、信号转导途径、蛋白质分解代谢过程、酰胺代谢过程和rna结合。利用C9orf72扩增患者的尸检脑组织并辅以细胞培养分析，我们评估了polyGA和valosin containing protein （VCP）之间的相互作用。这种相互作用的功能分析揭示了VCP与polyGA聚集体的隔离，改变了可溶性含缬草苷蛋白的水平。VCP也在自噬过程中起作用，与此一致，我们观察到表达polyGA的细胞的自噬改变。我们还观察到，在VCP缺失的细胞中，polyGA聚集体和p62的共定位发生了改变。综上所述，这些数据表明，polyGA聚集体对VCP的隔离有助于VCP功能的丧失，从而导致C9orf72扩张疾病中自噬过程的改变。利用BioID2接近标记，Božič等人鉴定了与C9orf72重复扩增相关的所有五种二肽重复蛋白的相互作用组。polyGA与VCP相互作用的功能分析表明，VCP被polyGA聚集体隔离，导致VCP功能的部分丧失。
Repeat expansions in the C9orf72 gene are a common cause of amyotrophic lateral sclerosis and frontotemporal lobar degeneration, two devastating neurodegenerative disorders. One of the proposed mechanisms of GGGGCC repeat expansion is their translation into non-canonical dipeptide repeats, which can then accumulate as aggregates and contribute to these pathologies. There are five different dipeptide repeat proteins (polyGA, polyGR, polyPR, polyPA and polyGP), some of which are known to be neurotoxic. In the present study, we used BioID2 proximity labelling to identify the interactomes of all five dipeptide repeat proteins consisting of 125 repeats each. We identified 113 interacting partners for polyGR, 90 for polyGA, 106 for polyPR, 25 for polyPA and 27 for polyGP. Gene Ontology enrichment analysis of the proteomic data revealed that these target interaction partners are involved in a variety of functions, including protein translation, signal transduction pathways, protein catabolic processes, amide metabolic processes and RNA-binding. Using autopsy brain tissue from patients with C9orf72 expansion complemented with cell culture analysis, we evaluated the interactions between polyGA and valosin containing protein (VCP). Functional analysis of this interaction revealed sequestration of VCP with polyGA aggregates, altering levels of soluble valosin-containing protein. VCP also functions in autophagy processes, and consistent with this, we observed altered autophagy in cells expressing polyGA. We also observed altered co-localization of polyGA aggregates and p62 in cells depleted of VCP. All together, these data suggest that sequestration of VCP with polyGA aggregates contributes to the loss of VCP function, and consequently to alterations in autophagy processes in C9orf72 expansion disorders. Using BioID2 proximity labelling, Božič et al. identify the interactomes of all five dipeptide repeat proteins associated with C9orf72 repeat expansion. Functional analysis of the interaction between polyGA and VCP reveals sequestration of VCP with polyGA aggregates, resulting in partial loss of VCP function.
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影响因子：
16.2
作者：
Ash PE;Bieniek KF;Gendron TF;Caulfield T;Lin WL;Dejesus-Hernandez M;van Blitterswijk MM;Jansen-West K;Paul JW 3rd;Rademakers R;Boylan KB;Dickson DW;Petrucelli L
通讯作者：
Petrucelli L
影响因子：
14.9
作者：
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Lempicki RA
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64.5
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Fernández-Busnadiego R
影响因子：
4.6
作者：
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通讯作者：
Neutzner, Albert
影响因子：
5.7
作者：
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