The domain of unknown function 4005 (DUF4005) in an Arabidopsis IQD protein functions in microtubule binding.

The domain of unknown function 4005 (DUF4005) in an Arabidopsis IQD protein functions in microtubule binding.
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拟南芥 IQD 蛋白中的未知功能域 4005 (DUF4005) 在微管结合中发挥作用

DOI:
10.1016/j.jbc.2021.100849
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发表时间:
2021-07
期刊:
The Journal of biological chemistry
影响因子:
--
通讯作者:
Liu X
Liu X
中科院分区:
其他
文献类型:
--
作者:
Li Y;Huang Y;Wen Y;Wang D;Liu H;Li Y;Zhao J;An L;Yu F;Liu X

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微管(MT)对内外信号的动态响应受到大量微管相关蛋白(MAPs)的调节。在高等植物中,许多植物特有的MAP在进化过程中出现,有利于它们的固着生活方式。IQ 67结构域(IQD)蛋白家族的一些成员已被证明是植物特异性MAP。然而,IQD蛋白和MT之间的相互作用机制仍然难以捉摸。在这里,我们证明了未知功能的结构域4005(DUF 4005)的拟南芥IQD家族蛋白ABS 6/AtIQD 16是一个新的MT结合结构域。共沉淀分析表明DUF 4005结构域在体外直接与MT结合。GFP标记的DUF 4005还修饰了体内测试的所有类型的MT阵列。此外,我们发现DUF 4005结构域内的15个氨基酸残基的保守延伸,其与人MAP Kif 18 A的C-末端MT结合结构域共享序列相似性,是与MT结合所需的。过表达DUF 4005结构域的转基因株系表现出一系列发育缺陷,包括在生物体水平上的螺旋生长和发育不良。在细胞水平上,DUF 4005过表达引起表皮铺装细胞和毛状体形态发生的缺陷,以及在黑暗生长的幼苗的下胚轴中的异常各向异性细胞伸长。这些数据表明ABS 6/AtIQD 16的DUF 4005结构域是一个新的MT结合结构域,其过表达扰乱植物中的MT稳态。我们的研究结果提供了新的见解MT-植物IQD蛋白的结合机制。
The dynamic responses of microtubules (MTs) to internal and external signals are modulated by a plethora of microtubule-associated proteins (MAPs). In higher plants, many plant-specific MAPs have emerged during evolution as advantageous to their sessile lifestyle. Some members of the IQ67 domain (IQD) protein family have been shown to be plant-specific MAPs. However, the mechanisms of interaction between IQD proteins and MTs remain elusive. Here we demonstrate that the domain of unknown function 4005 (DUF4005) of the Arabidopsis IQD family protein ABS6/AtIQD16 is a novel MT-binding domain. Cosedimentation assays showed that the DUF4005 domain binds directly to MTs in vitro. GFP-labeled DUF4005 also decorates all types of MT arrays tested in vivo. Furthermore, we showed that a conserved stretch of 15 amino acid residues within the DUF4005 domain, which shares sequence similarity with the C-terminal MT-binding domain of human MAP Kif18A, is required for the binding to MTs. Transgenic lines overexpressing the DUF4005 domain displayed a spectrum of developmental defects, including spiral growth and stunted growth at the organismal level. At the cellular level, DUF4005 overexpression caused defects in epidermal pavement cell and trichome morphogenesis, as well as abnormal anisotropic cell elongation in the hypocotyls of dark-grown seedlings. These data establish that the DUF4005 domain of ABS6/AtIQD16 is a new MT-binding domain, overexpression of which perturbs MT homeostasis in plants. Our findings provide new insights into the MT-binding mechanisms of plant IQD proteins.
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