DYF-5/MAK-dependent phosphorylation promotes ciliary tubulin unloading.
DYF-5/MAK-dependent phosphorylation promotes ciliary tubulin unloading.
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DYF-5/MAK 依赖性磷酸化促进纤毛微管蛋白卸载
DOI：
10.1073/pnas.2207134119
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发表时间：
2022-08-23
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
引起肌动蛋白和微管蛋白亚基显着动力学的分子机制一直是广泛研究的焦点。微管蛋白的严格调控的输入/输出对于纤毛发生特别重要，因为微管蛋白形成所有纤毛的轴丝“骨架”。与IFT-74/81亚基的相互作用有助于将微管蛋白装载到鞭毛内运输（IFT）机器上用于纤毛输入。我们发现，保守的DYF-5/MAK激酶定位在睫状体尖端磷酸化IFT-74，降低微管蛋白和IFT-74/81之间的结合亲和力六倍。在体内操作IFT-74的磷酸化状态改变纤毛长度。我们认为，DYF-5/MAK依赖性磷酸化促进微管蛋白在纤毛尖端卸载，并有助于微管蛋白动力学的位点特异性调节，以维持轴丝的完整性。纤毛是基于微管的细胞器，其为细胞运动提供动力并调节感觉和信号传导，并且纤毛结构和功能异常引起各种纤毛病。纤毛的形成和维持需要鞭毛内转运（IFT），在此期间，驱动蛋白-2家族马达蛋白将携带轴丝前体（如微管蛋白）的IFT颗粒运送到纤毛中。微管蛋白二聚体通过微管蛋白和IFT-74/81模块之间的相互作用加载到IFT机器;然而，很少有人知道微管蛋白到达睫状体尖端时是如何卸载的。在这里，我们表明，纤毛激酶DYF-5/MAK磷酸化多个网站内的微管蛋白结合模块的IFT-74，减少微管蛋白结合亲和力的IFT-74/81约六倍。IFT-74磷酸化的消融或组成性激活异常延长或缩短秀丽隐杆线虫神经元中的感觉纤毛。我们认为DYF-5/MAK依赖的磷酸化通过调节微管蛋白的卸载在纤毛发生中起着重要作用。
Molecular mechanisms that give rise to the remarkable dynamics of the actin and tubulin subunits have been a focus of extensive research. The tightly regulated import/export of tubulin is particularly important for ciliogenesis, given that tubulins form the axoneme “backbone” of all cilia. The interaction with the IFT-74/81 subunits helps load tubulin onto the intraflagellar transport (IFT) machinery for ciliary import. We found that the conserved DYF-5/MAK kinase localizing at the ciliary tip phosphorylates IFT-74, reducing the binding affinity between tubulin and IFT-74/81 sixfold. In vivo manipulation phosphorylation status of IFT-74 changed ciliary length. We propose that DYF-5/MAK–dependent phosphorylation promotes tubulin unloading at the ciliary tip and contributes to site-specific regulation of tubulin dynamics to maintain axonemal integrity. Cilia are microtubule-based organelles that power cell motility and regulate sensation and signaling, and abnormal ciliary structure and function cause various ciliopathies. Cilium formation and maintenance requires intraflagellar transport (IFT), during which the kinesin-2 family motor proteins ferry IFT particles carrying axonemal precursors such as tubulins into cilia. Tubulin dimers are loaded to IFT machinery through an interaction between tubulin and the IFT-74/81 module; however, little is known of how tubulins are unloaded when arriving at the ciliary tip. Here, we show that the ciliary kinase DYF-5/MAK phosphorylates multiple sites within the tubulin-binding module of IFT-74, reducing the tubulin-binding affinity of IFT-74/81 approximately sixfold. Ablation or constitutive activation of IFT-74 phosphorylation abnormally elongates or shortens sensory cilia in Caenorhabditis elegans neurons. We propose that DYF-5/MAK–dependent phosphorylation plays a fundamental role in ciliogenesis by regulating tubulin unloading.
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DOI：
10.1111/febs.14068
发表时间：
2017-09
期刊：
The FEBS journal
影响因子：
--
作者：
Prevo B;Scholey JM;Peterman EJG
通讯作者：
Peterman EJG
鞭毛内运输平衡外部双联微管的连续周转：对鞭毛长度控制的影响。
影响因子：
7.8
作者：
Marshall, W F;Rosenbaum, J L
通讯作者：
Rosenbaum, J L
影响因子：
64.8
作者：
Ou, GS;Blacque, OE;Scholey, JM
通讯作者：
Scholey, JM
影响因子：
9.8
作者：
Ozgul, Riza Koksal;Siemiatkowska, Anna M.;Corbo, Joseph C.
通讯作者：
Corbo, Joseph C.
影响因子：
7
作者：
Kinoshita, E;Kinoshita-Kikuta, E;Koike, T
通讯作者：
Koike, T