Departments of Pathology, University of California San Francisco, San Francisco, CA.
Department of Pathology, University of California San Diego, San Diego, CA.
J Cell Biol. 2022 Apr 4;221(4). doi: 10.1083/jcb.202010065. Epub 2022 Mar 10.
Missense mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson's disease (PD); however, pathways regulating LRRK2 subcellular localization, function, and turnover are not fully defined. We performed quantitative mass spectrometry-based interactome studies to identify 48 novel LRRK2 interactors, including the microtubule-associated E3 ubiquitin ligase TRIM1 (tripartite motif family 1). TRIM1 recruits LRRK2 to the microtubule cytoskeleton for ubiquitination and proteasomal degradation by binding LRRK2911-919, a nine amino acid segment within a flexible interdomain region (LRRK2853-981), which we designate the "regulatory loop" (RL). Phosphorylation of LRRK2 Ser910/Ser935 within LRRK2 RL influences LRRK2's association with cytoplasmic 14-3-3 versus microtubule-bound TRIM1. Association with TRIM1 modulates LRRK2's interaction with Rab29 and prevents upregulation of LRRK2 kinase activity by Rab29 in an E3-ligase-dependent manner. Finally, TRIM1 rescues neurite outgrowth deficits caused by PD-driving mutant LRRK2 G2019S. Our data suggest that TRIM1 is a critical regulator of LRRK2, controlling its degradation, localization, binding partners, kinase activity, and cytotoxicity.
LRRK2 中的错义突变是家族性帕金森病(PD)最常见的原因;然而,调节 LRRK2 亚细胞定位、功能和周转的途径尚未完全定义。我们进行了基于定量质谱的相互作用组学研究,以鉴定 48 种新的 LRRK2 相互作用蛋白,包括微管相关 E3 泛素连接酶 TRIM1(三基序家族 1)。TRIM1 通过结合 LRRK2911-919(位于柔性结构域间区(LRRK2853-981)内的九个氨基酸片段),将 LRRK2 募集到微管细胞骨架上进行泛素化和蛋白酶体降解,我们将其命名为“调节环”(RL)。LRRK2 RL 内的 Ser910/Ser935 磷酸化影响 LRRK2 与细胞质 14-3-3 与微管结合的 TRIM1 的关联。与 TRIM1 的关联调节 LRRK2 与 Rab29 的相互作用,并以 E3 连接酶依赖的方式防止 Rab29 上调 LRRK2 激酶活性。最后,TRIM1 挽救了由 PD 驱动的突变 LRRK2 G2019S 引起的神经突生长缺陷。我们的数据表明,TRIM1 是 LRRK2 的关键调节因子,控制其降解、定位、结合伙伴、激酶活性和细胞毒性。