Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Plant J. 2013 Mar;73(5):747-60. doi: 10.1111/tpj.12065. Epub 2013 Jan 17.
Rapid actin turnover is essential for numerous actin-based processes. However, how it is precisely regulated remains poorly understood. Actin-interacting protein 1 (AIP1) has been shown to be an important factor by acting coordinately with actin-depolymerizing factor (ADF)/cofilin in promoting actin depolymerization, the rate-limiting factor in actin turnover. However, the molecular mechanism by which AIP1 promotes actin turnover remains largely unknown in plants. Here, we provide a demonstration that AIP1 promotes actin turnover, which is required for optimal growth of rice plants. Specific down-regulation of OsAIP1 increased the level of filamentous actin and reduced actin turnover, whereas over-expression of OsAIP1 induced fragmentation and depolymerization of actin filaments and enhanced actin turnover. In vitro biochemical characterization showed that, although OsAIP1 alone does not affect actin dynamics, it enhances ADF-mediated actin depolymerization. It also caps the filament barbed end in the presence of ADF, but the capping activity is not required for their coordinated action. Real-time visualization of single filament dynamics showed that OsAIP1 enhanced ADF-mediated severing and dissociation of pointed end subunits. Consistent with this, the filament severing frequency and subunit off-rate were enhanced in OsAIP1 over-expressors but decreased in RNAi protoplasts. Importantly, OsAIP1 acts coordinately with ADF and profilin to induce massive net actin depolymerization, indicating that AIP1 plays a major role in the turnover of actin, which is required to optimize F-actin levels in plants.
快速肌动蛋白周转对于许多基于肌动蛋白的过程至关重要。然而,其确切的调节机制仍知之甚少。肌动蛋白相互作用蛋白 1(AIP1)已被证明是一个重要的因素,它与肌动蛋白解聚因子(ADF)/丝切蛋白协同作用,促进肌动蛋白解聚,这是肌动蛋白周转的限速步骤。然而,AIP1 促进肌动蛋白周转的分子机制在植物中仍知之甚少。在这里,我们提供了一个证据,表明 AIP1 促进了肌动蛋白的周转,这是水稻植物最佳生长所必需的。OsAIP1 的特异性下调增加了丝状肌动蛋白的水平并减少了肌动蛋白的周转,而 OsAIP1 的过表达诱导了肌动蛋白丝的片段化和解聚并增强了肌动蛋白的周转。体外生化特性分析表明,尽管 AIP1 本身并不影响肌动蛋白动力学,但它增强了 ADF 介导的肌动蛋白解聚。它还在 ADF 存在的情况下封闭肌动蛋白丝的加帽端,但加帽活性不是它们协调作用所必需的。单丝动力学的实时可视化显示,OsAIP1 增强了 ADF 介导的切断和尖端亚基的解离。与此一致的是,在 OsAIP1 过表达体中,丝的切断频率和亚基脱落率增加,而在 RNAi 原生质体中则降低。重要的是,OsAIP1 与 ADF 和 Profilin 协调作用,诱导大量净肌动蛋白解聚,这表明 AIP1 在肌动蛋白的周转中起着主要作用,这是优化植物中 F-肌动蛋白水平所必需的。