JNK-interacting protein 3 mediates the retrograde transport of activated c-Jun N-terminal kinase and lysosomes.
JNK-interacting protein 3 mediates the retrograde transport of activated c-Jun N-terminal kinase and lysosomes.
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DOI：
10.1371/journal.pgen.1003303
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发表时间：
2013
期刊：
影响因子：
4.5
通讯作者：
Nechiporuk AV
中科院分区：
文献类型：
--
作者：
Drerup CM;Nechiporuk AV
逆行轴突运输需要动力蛋白发动机和它的货物之间复杂的相互作用。是什么促成了这种互动在很大程度上是未知的。利用正向遗传学和一种新的体内成像方法，我们发现JNK相互作用蛋白3(JIP3)是基于动力蛋白的活化(磷酸化)c-Jun氨基末端激酶(JNK)和溶酶体逆行运输的直接中介。斑马鱼JIP3突变体(Jip3n17)表现出大的轴突末端肿胀，其中含有高水平的激活的JNK和溶酶体，但不包括其他逆行的货物，如晚期内体和自噬小体。利用体内轴突运输的分析，我们证明激活的JNK和溶酶体的终末积累是由于jip3n17中这些货物的逆行运动频率降低所致，而顺行运输在很大程度上没有受到影响。通过对缺乏JNK结合域的JIP3的抢救实验，我们进一步表明，JIP3-JNK相互作用的丧失是pJNK逆行运输缺陷的基础，从而导致轴突末端肿胀，而不是溶酶体积累。相反，我们的共转运分析表明，溶酶体积累是由于溶酶体与jip3n17中的动力蛋白轻中间链(动力蛋白辅助蛋白)失去了联系。因此，我们的结果表明，JIP3对于活性JNK和溶酶体这两种不同的货物的逆行运输是必需的。此外，我们的数据提供了强有力的证据，证明JIP3实际上是连接这些货物和动力蛋白的接头蛋白。为了形成和维持连接，神经元需要蛋白质和细胞器在神经元胞体和轴突终末之间的主动运输。抑制这种“轴突”运输被认为与神经退行性疾病有关。尽管这一过程很重要，但到目前为止，还没有一个脊椎动物模型系统可以在完整的动物身上研究轴突运输。我们的研究介绍了斑马鱼作为这样一个模型，并展示了它在分析轴突运输方面的能力。我们使用这个系统1)启动一个基因筛选以寻找轴突运输的新媒介；2)开发体内成像策略以实时显示完整动物的轴突运输；以及3)使用这些方法发现JNK相互作用蛋白3(JIP3)是从轴突终末到细胞体的两种货物运输所必需的(逆行运输)。在没有JIP3的情况下，这些货物堆积，轴突终末畸形，尽管其他货物的逆行运输是正常的。有趣的是，这些货物的异常定位与轴突疾病状态有关，但我们的工作是第一次确定它们从轴突终末运输所必需的特定适配蛋白。
Retrograde axonal transport requires an intricate interaction between the dynein motor and its cargo. What mediates this interaction is largely unknown. Using forward genetics and a novel in vivo imaging approach, we identified JNK-interacting protein 3 (Jip3) as a direct mediator of dynein-based retrograde transport of activated (phosphorylated) c-Jun N-terminal Kinase (JNK) and lysosomes. Zebrafish jip3 mutants (jip3nl7) displayed large axon terminal swellings that contained high levels of activated JNK and lysosomes, but not other retrograde cargos such as late endosomes and autophagosomes. Using in vivo analysis of axonal transport, we demonstrated that the terminal accumulations of activated JNK and lysosomes were due to a decreased frequency of retrograde movement of these cargos in jip3nl7, whereas anterograde transport was largely unaffected. Through rescue experiments with Jip3 engineered to lack the JNK binding domain and exogenous expression of constitutively active JNK, we further showed that loss of Jip3–JNK interaction underlies deficits in pJNK retrograde transport, which subsequently caused axon terminal swellings but not lysosome accumulation. Lysosome accumulation, rather, resulted from loss of lysosome association with dynein light intermediate chain (dynein accessory protein) in jip3nl7, as demonstrated by our co-transport analyses. Thus, our results demonstrate that Jip3 is necessary for the retrograde transport of two distinct cargos, active JNK and lysosomes. Furthermore, our data provide strong evidence that Jip3 in fact serves as an adapter protein linking these cargos to dynein. To form and maintain connections, neurons require the active transport of proteins and organelles between the neuronal cell body and axon terminals. Inhibition of this “axonal” transport has been linked to neurodegenerative diseases. Despite the importance of this process, to date there was no vertebrate model system where axonal transport could be studied in an intact animal. Our study introduces zebrafish as such a model and demonstrates its power for the analysis of axonal transport. We used this system to 1) initiate a genetic screen to find novel mediators of axonal transport; 2) develop in vivo imaging strategies to visualize axonal transport in real time in the intact animal; and 3) discover, using these methods, that JNK interacting protein 3 (Jip3) is required for the transport of two cargos, a kinase and lysosomes, from axon terminals to the cell body (retrograde transport). In the absence of Jip3, these cargos accumulate and axon terminals become dysmorphic, though the retrograde transport of other cargos is normal. Interestingly, abnormal localization of these cargos has been linked to axonal disease states, but our work is the first to identify a specific adapter protein necessary for their transport from axon terminals.
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影响因子：
64.5
作者：
Bowman, AB;Kamal, A;Goldstein, LSB
通讯作者：
Goldstein, LSB
影响因子：
9.2
作者：
Montagnac, Guillaume;Sibarita, Jean-Baptiste;Chavrier, Philippe
通讯作者：
Chavrier, Philippe
影响因子：
7.8
作者：
Cavalli, V;Kujala, P;Goldstein, LSB
通讯作者：
Goldstein, LSB
影响因子：
4.7
作者：
Miura, Eriko;Fukaya, Masahiro;Watanabe, Masahiko
通讯作者：
Watanabe, Masahiko
DOI：
10.1038/nrm2804
发表时间：
2009-12
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
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