Chemical screen of Arabidopsis zygote and proteomics in tobacco BY-2 cells identify general plant cell division inhibitors
Chemical screen of Arabidopsis zygote and proteomics in tobacco BY-2 cells identify general plant cell division inhibitors
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拟南芥受精卵的化学筛选和烟草 BY-2 细胞中的蛋白质组学鉴定了一般植物细胞分裂抑制剂
DOI：
10.1101/2022.04.28.489799
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Ueda Minako
中科院分区：
文献类型：
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作者：
Kimata Yusuke;Yamada Moe;Murata Takashi;Kuwata Keiko;Sato Ayato;Suzuki Takamasa;Kurihara Daisuke;Hasebe Mitsuyasu;Higashiyama Tetsuya;Ueda Minako
细胞分裂对生长发育至关重要，涉及纺锤体组装、染色体分离和细胞板形成等事件。在植物中，用于在所需时间控制这些事件的工具仍然很差，因为遗传方法由于高冗余性和致命性以及有害的副作用而无效。因此，我们筛选了影响细胞分裂的化合物，重点关注拟南芥拟合子，它们在母体胚珠中单独发育；无需延时观察，可以可靠地追踪细胞分裂。然后，我们使用烟草BY-2细胞进行活细胞成像和蛋白质组学鉴定鉴定化合物的靶事件。结果，我们分离到了两个化合物，PD-180970和PP2。PD-180970破坏微管（MT）组织，从而破坏核分离，可能是通过抑制MT相关蛋白（MAP70）。PP2影响了II类运动蛋白-12在膜质体出现部位的定位，并损害了细胞分裂。此外，这两种化学物质均对黄瓜（Cucumis sativus）和苔藓（Physcomitrium patens）等多种植物的生存力造成不可逆的损害。我们提出，基于拟南芥受精卵的化学筛选和以烟草BY-2细胞为重点的靶事件规范相结合，可以有效地识别新的工具和瞬时控制在不同植物物种中保守的特定细胞分裂事件。
Cell division is essential for growth and development and involves events such as spindle assembly, chromosome separation, and cell plate formation. In plants, the tools used to control these events at the desired time are still poor because the genetic approach is ineffective owing to a high redundancy and lethality, as well as harmful side effects. Accordingly, we screened cell division-affecting compounds, with a focus onArabidopsis thalianazygotes, which individually develop in maternal ovules; the cell division was reliably traceable without time-lapse observations. We then identified the target events of the identified compounds using tobacco BY-2 cells for live-cell imaging and proteomics. As a result, we isolated two compounds, PD-180970 and PP2. PD-180970 disrupts microtubule (MT) organization and, thus, nuclear separation, presumably by inhibiting MT-associated proteins (MAP70). PP2 affected class II Kinesin-12 localization at the phragmoplast emerging site and impaired cytokinesis. Moreover, neither chemical caused irreversible damage to viability but they were effective in multiple plant species such as cucumber (Cucumis sativus) and moss (Physcomitrium patens). We propose that the combination of chemical screening based on Arabidopsis zygotes and target event specification focusing on tobacco BY-2 cells can be used to effectively identify novel tools and transiently control specific cell division events that are conserved in diverse plant species.