BICD2, dynactin, and LIS1 cooperate in regulating dynein recruitment to cellular structures.
BICD2, dynactin, and LIS1 cooperate in regulating dynein recruitment to cellular structures.
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DOI：
10.1091/mbc.e12-03-0210
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发表时间：
2012-11
影响因子：
3.3
通讯作者：
Akhmanova A
中科院分区：
文献类型：
--
作者：
Splinter D;Razafsky DS;Schlager MA;Serra-Marques A;Grigoriev I;Demmers J;Keijzer N;Jiang K;Poser I;Hyman AA;Hoogenraad CC;King SJ;Akhmanova A
本研究剖析了动力蛋白和动力蛋白的招募，以货物保守的马达适配器BICD2。结果表明，dynein、dynactin和BICD2在体内和体外形成一个三重复合物。通过在活细胞中直接观察动力蛋白对该复合物性质的研究表明，bicd2诱导的动力蛋白运输需要LIS1。细胞质动力蛋白是主要的微管负端定向细胞马达。大多数动力蛋白活动需要动力蛋白，但调节货物依赖性动力蛋白-动力蛋白相互作用的机制尚不清楚。在这项研究中，我们关注的是动力蛋白-动力蛋白通过保守的运动适配器Bicaudal D2 （BICD2）向货物的补充。我们发现，在BICD2介导的货物靶向中，动力蛋白和动动蛋白相互依赖，BICD2 n端（BICD2- n）在体内和体外都强烈促进动力蛋白和动动蛋白之间稳定的相互作用。活细胞中动力蛋白的直接可视化表明，bicd2 - n -动力蛋白-动力蛋白复合物本身不能与货物或微管相互作用。然而，BICD2-N系在不同的膜上促进了它们的微管负端定向运动。我们进一步证明，由BICD2-N的膜系结诱导的动力蛋白介导的运输需要LIS1，并且LIS1有助于微管+端和bicd2阳性细胞结构的动力蛋白积累。我们的研究结果表明，动力蛋白向货物的招募需要多个动力蛋白辅助因子的协同作用。
This study dissects the recruitment of dynein and dynactin to cargo by a conserved motor adaptor BICD2. It is shown that dynein, dynactin, and BICD2 form a triple complex in vitro and in vivo. Investigation of the properties of this complex by direct visualization of dynein in live cells shows that BICD2-induced dynein transport requires LIS1. Cytoplasmic dynein is the major microtubule minus-end–directed cellular motor. Most dynein activities require dynactin, but the mechanisms regulating cargo-dependent dynein–dynactin interaction are poorly understood. In this study, we focus on dynein–dynactin recruitment to cargo by the conserved motor adaptor Bicaudal D2 (BICD2). We show that dynein and dynactin depend on each other for BICD2-mediated targeting to cargo and that BICD2 N-terminus (BICD2-N) strongly promotes stable interaction between dynein and dynactin both in vitro and in vivo. Direct visualization of dynein in live cells indicates that by itself the triple BICD2-N–dynein–dynactin complex is unable to interact with either cargo or microtubules. However, tethering of BICD2-N to different membranes promotes their microtubule minus-end–directed motility. We further show that LIS1 is required for dynein-mediated transport induced by membrane tethering of BICD2-N and that LIS1 contributes to dynein accumulation at microtubule plus ends and BICD2-positive cellular structures. Our results demonstrate that dynein recruitment to cargo requires concerted action of multiple dynein cofactors.