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A novel strategy to visualize vesicle-bound kinesins reveals the diversity of kinesin-mediated transport.
Traffic ( IF 3.6 ) Pub Date : 2019-10-02 , DOI: 10.1111/tra.12692
Rui Yang 1, 2 , Zoe Bostick 3 , Alex Garbouchian 3 , Julie Luisi 1 , Gary Banker 1 , Marvin Bentley 3
Affiliation  

In mammals, 15 to 20 kinesins are thought to mediate vesicle transport. Little is known about the identity of vesicles moved by each kinesin or the functional significance of such diversity. To characterize the transport mediated by different kinesins, we developed a novel strategy to visualize vesicle-bound kinesins in living cells. We applied this method to cultured neurons and systematically determined the localization and transport parameters of vesicles labeled by different members of the Kinesin-1, -2, and -3 families. We observed vesicle labeling with nearly all kinesins. Only six kinesins bound vesicles that undergo long-range transport in neurons. Of these, three had an axonal bias (KIF5B, KIF5C and KIF13B), two were unbiased (KIF1A and KIF1Bβ), and one transported only in dendrites (KIF13A). Overall, the trafficking of vesicle-bound kinesins to axons or dendrites did not correspond to their motor domain preference, suggesting that on-vesicle regulation is crucial for kinesin targeting. Surprisingly, several kinesins were associated with populations of somatodendritic vesicles that underwent little long-range transport. This assay should be broadly applicable for investigating kinesin function in many cell types.

中文翻译:

一种可视化囊泡结合驱动蛋白的新策略揭示了驱动蛋白介导的运输的多样性。

在哺乳动物中,15 至 20 个驱动蛋白被认为介导囊泡运输。对于每种驱动蛋白移动的囊泡的身份或这种多样性的功能意义知之甚少。为了表征不同驱动蛋白介导的运输,我们开发了一种新策略来可视化活细胞中囊泡结合的驱动蛋白。我们将此方法应用于培养的神经元,并系统地确定了由驱动蛋白-1、-2和-3家族不同成员标记的囊泡的定位和运输参数。我们观察到几乎所有驱动蛋白的囊泡标记。只有六种驱动蛋白结合在神经元中进行长程运输的囊泡。其中,三种具有轴突偏向(KIF5B、KIF5C 和 KIF13B),两种无偏向(KIF1A 和 KIF1Bβ),一种仅在树突中运输(KIF13A)。总体而言,囊泡结合的驱动蛋白向轴突或树突的运输与它们的运动域偏好并不相符,这表明囊泡上的调节对于驱动蛋白靶向至关重要。令人惊讶的是,几种驱动蛋白与几乎不进行长程运输的体细胞树突囊泡群体相关。该测定应广泛适用于研究许多细胞类型中的驱动蛋白功能。
更新日期:2019-10-02
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