Dos Passos Patricia M, Hemamali Erandika H, Mamede Lohany D, Hayes Lindsey R, Ayala Yuna M
Edward Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, Missouri, United States of America.
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
PLoS Biol. 2024 Feb 29;22(2):e3002527. doi: 10.1371/journal.pbio.3002527. eCollection 2024 Feb.
TDP-43 is an essential RNA-binding protein strongly implicated in the pathogenesis of neurodegenerative disorders characterized by cytoplasmic aggregates and loss of nuclear TDP-43. The protein shuttles between nucleus and cytoplasm, yet maintaining predominantly nuclear TDP-43 localization is important for TDP-43 function and for inhibiting cytoplasmic aggregation. We previously demonstrated that specific RNA binding mediates TDP-43 self-assembly and biomolecular condensation, requiring multivalent interactions via N- and C-terminal domains. Here, we show that these complexes play a key role in TDP-43 nuclear retention. TDP-43 forms macromolecular complexes with a wide range of size distribution in cells and we find that defects in RNA binding or inter-domain interactions, including phase separation, impair the assembly of the largest species. Our findings suggest that recruitment into these macromolecular complexes prevents cytoplasmic egress of TDP-43 in a size-dependent manner. Our observations uncover fundamental mechanisms controlling TDP-43 cellular homeostasis, whereby regulation of RNA-mediated self-assembly modulates TDP-43 nucleocytoplasmic distribution. Moreover, these findings highlight pathways that may be implicated in TDP-43 proteinopathies and identify potential therapeutic targets.
TDP-43是一种重要的RNA结合蛋白,与以细胞质聚集体和核TDP-43缺失为特征的神经退行性疾病的发病机制密切相关。该蛋白在细胞核和细胞质之间穿梭,但维持主要的核TDP-43定位对于TDP-43的功能以及抑制细胞质聚集很重要。我们之前证明,特定的RNA结合介导TDP-43的自组装和生物分子凝聚,这需要通过N端和C端结构域进行多价相互作用。在这里,我们表明这些复合物在TDP-43的核保留中起关键作用。TDP-43在细胞中形成具有广泛大小分布的大分子复合物,我们发现RNA结合或结构域间相互作用(包括相分离)的缺陷会损害最大复合物的组装。我们的研究结果表明,被招募到这些大分子复合物中以大小依赖的方式阻止了TDP-43的细胞质外流。我们的观察结果揭示了控制TDP-43细胞内稳态的基本机制,即RNA介导的自组装调节调节TDP-43的核质分布。此外,这些发现突出了可能与TDP-43蛋白病有关的途径,并确定了潜在的治疗靶点。