New function of the proline rich domain in dynamin-2 to negatively regulate its interaction with microtubules in mammalian cells
New function of the proline rich domain in dynamin-2 to negatively regulate its interaction with microtubules in mammalian cells
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DOI：
10.1016/j.yexcr.2009.01.025
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发表时间：
2009-04-15
影响因子：
3.7
通讯作者：
Hosoya, Hiroshi
中科院分区：
文献类型：
--
作者：
Hamao, Kozue;Morita, Makiko;Hosoya, Hiroshi
微管重组对于许多细胞功能如细胞迁移、细胞极性和细胞分裂是必需的。发动蛋白最初被鉴定为微管结合蛋白。先前的有限消化实验表明发动蛋白C端100个氨基酸的富含脯氨酸的结构域（PRD）负责微管的体外结合。然而，由于在有丝分裂过程中，动力蛋白仅在纺锤体中间区而不是在间期细胞中沿沿着微管观察到明显的定位，因此动力蛋白在体内如何与微管相互作用仍不清楚。在这里，我们报告GFP-发动蛋白-2-（1-786），一个截短的突变体，缺乏PRO的C-末端部分，定位于沿着微管在间期HeLa细胞。GFP-发动蛋白-2-野生型（WT）和GFP-发动蛋白-2-（1-745），一种被进一步截短以去除整个PRD的构建体，定位在离散的点状结构中，但不沿着微管。这些数据表明，N-末端（残基746-786）而不是整个PRO是发动蛋白-2与细胞中微管相互作用所必需的，PRO的C-末端（787-870）负调控这种相互作用。表达GFP-发动蛋白-2-（1-786）的细胞中的微管针对暴露于冷而稳定。这些结果提供了第一个证据，在培养的哺乳动物细胞中的微管调节动力蛋白-2的相互作用。(C)2009爱思唯尔公司All rights reserved.
Microtubule reorganization is necessary for many cellular functions such as cell migration, cell polarity and cell division. Dynamin was originally identified as a microtubule-binding protein. Previous limited digestion experiment revealed that C-terminal 100-amino acids proline rich domain (PRD) of dynamin is responsible for microtubule binding in vitro. However, as obvious localization of dynamin along microtubules is only observed at the spindle midzone during mitosis but not in interphase cells, it remains unclear how dynamin interacts with microtubules in vivo. Here, we report that GFP-dynamin-2-(1-786), a truncated mutant lacking a C-terminal portion of the PRO, localized along rnicrotubules in interphase HeLa cells. GFP-dynamin-2-wild type (WT) and GFP-dynamin-2-(1-745), a construct that was further truncated to remove the entire PRD, localized in discrete punctuate structures but not along rnicrotubules. These data suggest that the N-terminal (residues 746-786) but not the entire PRO is necessary for the interaction of dynamin-2 with microtubules in the cell and that the C-terminus of PRO (787-870) negatively regulate this interaction. Microtubules in cells expressing GFP-dynamin-2-(1-786) were stabilized against exposure to cold. These results provide a first evidence for a regulated interaction of dynamin-2 with microtubules in cultured mammalian cells. (C) 2009 Elsevier Inc. All rights reserved.