Cell cycle dependent dynamics of a plant intermediate filament motif protein with intracellular localization related to microtubules
Cell cycle dependent dynamics of a plant intermediate filament motif protein with intracellular localization related to microtubules
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具有与微管相关的细胞内定位的植物中间丝基序蛋白的细胞周期依赖性动力学
DOI：
10.1007/s00709-020-01512-1
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发表时间：
2020
期刊：
影响因子：
2.9
通讯作者：
Tsuyoshi Kaneta
中科院分区：
文献类型：
--
作者：
Hikaru Utsunomiya;Masayuki Fujita;Fumio Naito;Tsuyoshi Kaneta
虽然生物化学和免疫学认为中间丝存在于植物细胞中，但与形成这些结构的蛋白质相关的分子遗传学研究很少。在本研究中，拟南芥AT3G05270被选为植物细胞中构成IF蛋白的候选基因。AT3G05270编码的蛋白质具有一个大的α-螺旋以及维持IF结构所必需的IF蛋白基序。此外，该蛋白与GFP融合后的荧光信号在植物细胞中呈现细胞骨架样的丝状结构。因此，我们将AT3G05270编码的蛋白质命名为中间丝基序蛋白1(IFMoP1)。在表达IFMoP1-GFP的烟草BY-2细胞中检测了由IFMoP1组成的结构及其定位，该细胞的细胞周期用DNA合成抑制剂aphidiclin和微管干扰剂丙氨酰胺同步。IFMoP1-GFP信号出现在有丝分裂期的纺锤体和成纤维细胞上。此外，IFMoP1-GFP组成的细胞骨架样丝状结构的细胞频率随着完成细胞分裂的细胞的增加而增加，几小时后下降。根据IFMoP1与微管在细胞内定位的关系，IFMoP1组成的丝状结构在间期独立于微管存在。在活细胞中，这些丝状结构与细胞核一起移动。IFMoP1在有丝分裂过程中与纺锤形微管和成纤维细胞微管共定位，间期与部分皮质微管共定位。
Although intermediate filaments (IFs) are biochemically and immunologically suggested to exist in plant cells, there are few molecular genetic studies related to the proteins that form these structures. In this study,ArabidopsisAT3G05270 was selected as a candidate gene for a protein constituting IF in plant cells. The protein encoded by AT3G05270 has a large α-helix as well as the IF protein motif indispensable for maintaining the structures of IF. Moreover, fluorescence signals of this protein fused with GFP exhibited cytoskeleton-like filamentous structures in plant cells. Thus, we named the protein encoded by AT3G05270 as Intermediate Filament Motif Protein 1 (IFMoP1). The structures composed of IFMoP1 and their localizations were examined in IFMoP1-GFP-expressing tobacco BY-2 cells whose cell cycle was synchronized using aphidicolin, a DNA synthesis inhibitor, and propyzamide, a microtubule-disrupting agent. The IFMoP1-GFP signals were present at the spindles and phragmoplasts in the mitotic phase. In addition, the frequency of cells with cytoskeleton-like filamentous structures composed of IFMoP1-GFP increased with the increase in cells that completed cell division, and then decreased after several hours. In terms of the relationship in intracellular localization between IFMoP1 and microtubules, the filamentous structures composed of IFMoP1 were present independently of microtubules during interphase. In living cells, these filamentous structures moved along with the nucleus. IFMoP1 co-localized with spindle and phragmoplast microtubules during mitosis, as well as with a part of the cortical microtubules in interphase.