LRRK2 transport is regulated by its novel interacting partner Rab32.
LRRK2 transport is regulated by its novel interacting partner Rab32.
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DOI：
10.1371/journal.pone.0111632
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Barnekow A
中科院分区：
文献类型：
--
作者：
Waschbüsch D;Michels H;Strassheim S;Ossendorf E;Kessler D;Gloeckner CJ;Barnekow A
富含亮氨酸重复序列激酶2（LRRK 2）是一种与帕金森病（PD）相关的多结构域280 kDa蛋白。突变，特别是在GT3和LRRK 2的激酶结构域中的突变是遗传性PD的最常见原因，并且也在散发形式的PD中发现。虽然LRRK 2的细胞功能在很大程度上是未知的，但越来越多的证据表明，这些突变由于自噬功能障碍和线粒体损伤而导致细胞死亡。在这里，我们证明了一种新的机制LRRK 2的结合和运输，其中涉及小GTP酶Rab 32和Rab 38。Rab 32及其最接近的同源物Rab 38已知组织trans-Golgi网络和运输黑素生成中的关键酶，而它们在非黑素生成细胞中的功能仍然没有很好地理解。细胞过程如自噬、线粒体动力学、吞噬作用或脑中的炎症过程先前已与Rab 32相关。在这里，我们证明了Rab 32和Rab 38，但没有其他GTK测试，直接与LRRK 2相互作用。GFP-Trap分析证实Rab 32与内源性LRRK 2的相互作用。在酵母双杂交实验中，我们确定了一个预测的卷曲螺旋基序包含区域内的氨基端的LRRK 2作为可能的相互作用域。荧光显微镜显示Rab 32和LRRK 2在再循环内体和转运囊泡中的共定位，而Rab 32的组成型活性突变体的过表达导致Rab 7/9阳性核周晚期内体/MVB的共定位增加。亚细胞分级实验支持Rab 32在LRRK 2晚期内体转运和细胞分选中的新作用。因此，Rab 32可能调节LRRK 2的生理功能。
Leucine-rich repeat kinase 2 (LRRK2) is a multi-domain 280 kDa protein that is linked to Parkinson's disease (PD). Mutations especially in the GTPase and kinase domains of LRRK2 are the most common causes of heritable PD and are also found in sporadic forms of PD. Although the cellular function of LRRK2 is largely unknown there is increasing evidence that these mutations cause cell death due to autophagic dysfunction and mitochondrial damage. Here, we demonstrate a novel mechanism of LRRK2 binding and transport, which involves the small GTPases Rab32 and Rab38. Rab32 and its closest homologue Rab38 are known to organize the trans-Golgi network and transport of key enzymes in melanogenesis, whereas their function in non-melanogenic cells is still not well understood. Cellular processes such as autophagy, mitochondrial dynamics, phagocytosis or inflammatory processes in the brain have previously been linked to Rab32. Here, we demonstrate that Rab32 and Rab38, but no other GTPase tested, directly interact with LRRK2. GFP-Trap analyses confirmed the interaction of Rab32 with the endogenous LRRK2. In yeast two-hybrid experiments we identified a predicted coiled-coil motif containing region within the aminoterminus of LRRK2 as the possible interacting domain. Fluorescence microscopy demonstrated a co-localization of Rab32 and LRRK2 at recycling endosomes and transport vesicles, while overexpression of a constitutively active mutant of Rab32 led to an increased co-localization with Rab7/9 positive perinuclear late endosomes/MVBs. Subcellular fractionation experiments supported the novel role of Rab32 in LRRK2 late endosomal transport and sorting in the cell. Thus, Rab32 may regulate the physiological functions of LRRK2.
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DOI：
10.1073/pnas.1318306111
发表时间：
2014-02-18
影响因子：
11.1
作者：
Beilina, Alexandria;Rudenko, Iakov N.;Cookson, Mark R.
通讯作者：
Cookson, Mark R.
影响因子：
3
作者：
Gillardon, Frank;Kremmer, Elisabeth;Gloeckner, Christian J.
通讯作者：
Gloeckner, Christian J.
影响因子：
11.2
作者：
Biskup, Saskia;Moore, Darren J.;Dawson, Valina L.
通讯作者：
Dawson, Valina L.
DOI：
10.1083/jcb.200204081
发表时间：
2002-08-19
期刊：
The Journal of cell biology
影响因子：
--
作者：
Alto NM;Soderling J;Scott JD
通讯作者：
Scott JD
影响因子：
3.5
作者：
Gloeckner, CJ;Kinkl, N;Ueffing, M
通讯作者：
Ueffing, M