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Arabidopsis RIC1 Severs Actin Filaments at the Apex to Regulate Pollen Tube Growth

机译:拟南芥RIC1切断了肌动蛋白丝调节花粉管的生长

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摘要

Pollen tubes deliver sperms to the ovule for fertilization via tip growth. The rapid turnover of F-actin in pollen tube tips plays an important role in this process. In this study, we demonstrate that Arabidopsis thaliana RIC1, a member of the ROP-interactive CRIB motif-containing protein family, regulates pollen tube growth via its F-actin severing activity. Knockout of RIC1 enhanced pollen tube elongation, while overexpression of RIC1 dramatically reduced tube growth. Pharmacological analysis indicated that RIC1 affected F-actin dynamics in pollen tubes. In vitro biochemical assays revealed that RIC1 directly bound and severed F-actin in the presence of Ca2+ in addition to interfering with F-actin turnover by capping F-actin at the barbed ends. In vivo, RIC1 localized primarily to the apical plasma membrane () of pollen tubes. The level of RIC1 at the apical oscillated during pollen tube growth. The frequency of F-actin severing at the apex was notably decreased in ric1-1 pollen tubes but was increased in pollen tubes overexpressing RIC1. We propose that RIC1 regulates F-actin dynamics at the apical as well as the cytosol by severing F-actin and capping the barbed ends in the cytoplasm, establishing a novel mechanism that underlies the regulation of pollen tube growth.
机译:花粉管通过顶端生长将精子传递到胚珠进行受精。花粉管尖端中F-肌动蛋白的快速周转在此过程中起重要作用。在这项研究中,我们证明了拟南芥RIC1,它是含有ROP相互作用的CRIB基序的蛋白质家族的成员,通过其F-肌动蛋白切断活性来调节花粉管的生长。 RIC1的敲除增强了花粉管的伸长,而RIC1的过表达显着降低了花粉管的生长。药理分析表明,RIC1影响花粉管中的F-肌动蛋白动力学。体外生化分析显示,RIC1在Ca 2 + 存在下直接结合并切断F-肌动蛋白,此外还通过在倒钩末端封端F-肌动蛋白来干扰F-肌动蛋白更新。在体内,RIC1主要定位在花粉管的顶质膜上。在花粉管生长过程中,顶端的RIC1水平振荡。在ric1-1花粉管中,F-肌动蛋白在顶点处切断的频率显着降低,而在过表达RIC1的花粉管中则增加。我们提出,RIC1通过切断F-actin并封盖细胞质的倒刺末端来调节顶端以及细胞质中的F-actin动力学,从而建立了一种新的机制来支撑花粉管的生长。

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