Molecular Cell Biology, Botanical Institute, and Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), Kaiserstrasse 2, D-76131 Karlsruhe, Germany.
New Phytol. 2012 Feb;193(3):576-589. doi: 10.1111/j.1469-8137.2011.03944.x. Epub 2011 Nov 10.
• Kinesins with a calponin homology domain (KCHs) have been identified recently as a plant-specific subgroup of the kinesin-14 family and are suspected to act as microtubule-actin filament cross-linkers. The cellular function, however, has remained elusive. • In order to address the function of KCHs, we isolated NtKCH, a novel KCH homologue from tobacco BY-2 cells. Following synchronization, NtKCH transcripts were shown to be abundant during mitosis, whereas, during interphase, expression was low. • Using fluorescent-tagged cell lines and immunolabelling techniques, the localization of tobacco KCH was found to differ depending on the cell cycle. During interphase, NtKCH mainly associated with cortical microtubules, whereas a subfraction also co-localized with perinuclear actin cables. In dividing cells, NtKCH accumulated at the pre-prophase band and at the phragmoplast. However, it remained absent from spindle microtubules, but, instead, concentrated at two agglomerations in proximity to the two cell poles. • This work develops a detailed model for the dual localization and function of NtKCH during cell division vs cell expansion. This model implies two dynamic states of KCHs that differ with regard to actin interaction. This allows the modulation of force generation by KCH in a cell cycle-dependent capture mechanism.
• 最近,人们发现具有钙调蛋白同源结构域的驱动蛋白(KCHs)是驱动蛋白-14 家族中的一个植物特异性亚群,据推测其作用是微管-肌动蛋白丝交联剂。然而,其细胞功能仍然难以捉摸。• 为了研究 KCH 的功能,我们从烟草 BY-2 细胞中分离出一种新的 KCH 同源物 NtKCH。在同步化后,发现 NtKCH 转录本在有丝分裂期间丰度很高,而在间期间,表达水平较低。• 使用荧光标记的细胞系和免疫标记技术,发现烟草 KCH 的定位因细胞周期而异。在间期间,NtKCH 主要与皮质微管相关,而一小部分也与核周肌动蛋白电缆共定位。在分裂的细胞中,NtKCH 积累在前中期带和成膜体中。然而,它在纺锤体微管中缺失,但在两个细胞极附近的两个聚集处浓缩。• 这项工作为 NtKCH 在细胞分裂与细胞扩张过程中的双重定位和功能建立了一个详细的模型。该模型暗示 KCHs 有两种动态状态,它们在与肌动蛋白的相互作用方面存在差异。这允许 KCH 通过细胞周期依赖性捕获机制来调节力的产生。