Distinctive roles of Rac1 and Rab29 in LRRK2 mediated membrane trafficking and neurite outgrowth.
Distinctive roles of Rac1 and Rab29 in LRRK2 mediated membrane trafficking and neurite outgrowth.
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Rac1 和 Rab29 在 LRRK2 介导的膜运输和神经突生长中的独特作用
DOI：
10.7555/jbr.31.20170039
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发表时间：
2018-03-26
影响因子：
2.3
通讯作者：
Liu YJ
中科院分区：
文献类型：
--
作者：
Feng M;Hu X;Li N;Hu F;Chang F;Xu HF;Liu YJ
帕金森病（Parkinson's disease，PD）相关的富含亮氨酸重复序列激酶2（leucine-rich repeat kinase 2，LRRK 2）突变体对多种亚细胞过程具有致病作用，两种小GTP酶Rac 1和Rab 29被认为是参与LRRK 2信号转导的可能下游效应子，但其具体机制尚不清楚。在本研究中，我们使用生物化学和细胞生物学方法来研究两种GTP酶是否与LRRK 2相互作用，从而在LRRK 2介导的发病机制中发挥不同的功能，我们发现Rac 1和Rab 29特异性地与LRRK 2相互作用，对Rab 29具有更高的亲和力，并且在功能域结合中具有不同的偏好。突变Rab 29而不是Rac 1改变了货物蛋白阳离子非依赖性甘露糖-6-磷酸受体（CI-M6 PR）的内体到TGN的逆行运输及其稳定性。另一方面，过量表达的野生型Rab 29而不是Rac 1拯救了由致病突变体LRRK 2G 2019 S诱导的改变的逆行膜运输。此外，Rac 1和Rab 29都可以挽救LRRK 2G 2019 S诱导的分化的SH-SY 5 Y细胞的神经突起缩短。我们的研究强烈表明Rac 1和Rab 29作为LRRK 2信号通路的下游效应子参与了不同的功能。
Parkinson’s disease (PD) associated leucine-rich repeat kinase 2 (LRRK2) mutants have shown pathogenic effects on variety of subcellular processes.Two small GTPases Rac1 and Rab29 have been indicated as possible downstream effectors participating in LRRK2 signaling but their detail mechanisms remain unclear. In this study, we have used biochemical and cell biology approaches to address whether two GTPases interact with LRRK2 and hence function differently in LRRK2 mediated pathogenesis.Here we show thatRac1 and Rab29 specifically interact with LRRK2with higher affinity for Rab29and with different preference in functional domain binding. Mutant Rab29 but not Rac1 alters theendosome-to-TGN retrograde trafficking of a cargo protein cation-independent mannose-6-phosphate receptor (CI-M6PR) and its stability. On the other hand, overexpressedwild type Rab29 but not Rac1 rescue the altered retrograde membrane trafficking induced by the pathogenic mutant LRRK2G2019S. Furthermore, both Rac1 and Rab29 can rescue the neurite shortening in differentiated SH-SY5Y cells induced by LRRK2G2019S. Our study strongly suggests that Rac1 and Rab29 are involved in the distinct functions as downstream effe ctors in LRRK2 signaling pathways.