Fibril treatment changes protein interactions of tau and α-synuclein in human neurons.
Fibril treatment changes protein interactions of tau and α-synuclein in human neurons.
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DOI：
10.1016/j.jbc.2023.102888
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发表时间：
2023-03
影响因子：
4.8
通讯作者：
Carlson, George A.
中科院分区：
文献类型：
--
作者：
Griffin, Tagan A.;Schnier, Paul D.;Cleveland, Elisa M.;Newberry, Robert W.;Becker, Julia;Carlson, George A.
在几种神经退行性疾病中，神经元蛋白tau和α-突触核蛋白采用能够在细胞内和细胞间复制的聚集倾向构象。为了更好地理解这些构象变化如何驱动神经病理学，我们比较了在存在或不存在重组原纤维种子的情况下tau和α-突触核蛋白的相互作用体。将具有可诱导的神经生成素-2转基因的人胚胎干细胞分化为表达（1）WT 0 N4 R tau、（2）突变型（P301 L）0 N4 R tau、（3）WT α-突触核蛋白或（4）突变型（A53 T）α-突触核蛋白的神经元能神经元，每种神经元都与混杂生物素连接酶（BioID 2）基因融合。表达未融合BioID 2的神经元作为对照。在用原纤维或PBS处理后，用生物素原位标记相互作用的蛋白质，并通过串联质量标签标记使用质谱法进行定量。通过比较突变体与WT神经元以及原纤维与PBS处理的神经元中的相互作用，我们观察到可能与疾病进展相关的蛋白质相互作用的变化。我们确定了45个共同的相互作用因子，表明tau和α-突触核蛋白在一些相同的通路中起作用。共享相互作用的潜在位点包括微管、Wnt信号复合物和RNA颗粒。原纤维处理后，生理相互作用降低，而其他相互作用，包括tau和14-3-3 η之间的相互作用增加。我们证实，已知在神经退行性变期间与蛋白质聚集体共定位的14-3-3蛋白可以在体外促进或抑制tau聚集，具体取决于14-3-3亚型和tau序列的特定组合。
In several neurodegenerative disorders, the neuronal proteins tau and α-synuclein adopt aggregation-prone conformations capable of replicating within and between cells. To better understand how these conformational changes drive neuropathology, we compared the interactomes of tau and α-synuclein in the presence or the absence of recombinant fibril seeds. Human embryonic stem cells with an inducible neurogenin-2 transgene were differentiated into glutamatergic neurons expressing (1) WT 0N4R tau, (2) mutant (P301L) 0N4R tau, (3) WT α-synuclein, or (4) mutant (A53T) α-synuclein, each genetically fused to a promiscuous biotin ligase (BioID2). Neurons expressing unfused BioID2 served as controls. After treatment with fibrils or PBS, interacting proteins were labeled with biotin in situ and quantified using mass spectrometry via tandem mass tag labeling. By comparing interactions in mutant versus WT neurons and in fibril- versus PBS-treated neurons, we observed changes in protein interactions that are likely relevant to disease progression. We identified 45 shared interactors, suggesting that tau and α-synuclein function within some of the same pathways. Potential loci of shared interactions include microtubules, Wnt signaling complexes, and RNA granules. Following fibril treatment, physiological interactions decreased, whereas other interactions, including those between tau and 14-3-3 η, increased. We confirmed that 14-3-3 proteins, which are known to colocalize with protein aggregates during neurodegeneration, can promote or inhibit tau aggregation in vitro depending on the specific combination of 14-3-3 isoform and tau sequence.
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