Avisar Dror, Abu-Abied Mohamad, Belausov Eduard, Sadot Einat, Hawes Chris, Sparkes Imogen A
Institute of Plant Sciences, Volcani Center, Bet-Dagan 50250, Israel.
Plant Physiol. 2009 Jun;150(2):700-9. doi: 10.1104/pp.109.136853. Epub 2009 Apr 15.
Gene families with multiple members are predicted to have individuals with overlapping functions. We examined all of the Arabidopsis (Arabidopsis thaliana) myosin family members for their involvement in Golgi and other organelle motility. Truncated fragments of all 17 annotated Arabidopsis myosins containing either the IQ tail or tail domains only were fused to fluorescent markers and coexpressed with a Golgi marker in two different plants. We tracked and calculated Golgi body displacement rate in the presence of all myosin truncations and found that tail fragments of myosins MYA1, MYA2, XI-C, XI-E, XI-I, and XI-K were the best inhibitors of Golgi body movement in the two plants. Tail fragments of myosins XI-B, XI-F, XI-H, and ATM1 had an inhibitory effect on Golgi bodies only in Nicotiana tabacum, while tail fragments of myosins XI-G and ATM2 had a slight effect on Golgi body motility only in Nicotiana benthamiana. The best myosin inhibitors of Golgi body motility were able to arrest mitochondrial movement too. No exclusive colocalization was found between these myosins and Golgi bodies in our system, although the excess of cytosolic signal observed could mask myosin molecules bound to the surface of the organelle. From the preserved actin filaments found in the presence of enhanced green fluorescent protein fusions of truncated myosins and the motility of myosin punctae, we conclude that global arrest of actomyosin-derived cytoplasmic streaming had not occurred. Taken together, our data suggest that the above myosins are involved, directly or indirectly, in the movement of Golgi and mitochondria in plant cells.
预计具有多个成员的基因家族会有功能重叠的个体。我们研究了拟南芥(Arabidopsis thaliana)中所有肌球蛋白家族成员在高尔基体及其他细胞器运动中的作用。将17个已注释的拟南芥肌球蛋白仅包含IQ尾巴或尾巴结构域的截短片段与荧光标记融合,并在两种不同的植物中与高尔基体标记共表达。我们追踪并计算了在所有肌球蛋白截短情况下高尔基体的位移速率,发现肌球蛋白MYA1、MYA2、XI-C、XI-E、XI-I和XI-K的尾巴片段是两种植物中高尔基体运动的最佳抑制剂。肌球蛋白XI-B、XI-F、XI-H和ATM1的尾巴片段仅在烟草中对高尔基体有抑制作用,而肌球蛋白XI-G和ATM2的尾巴片段仅在本氏烟草中对高尔基体运动有轻微影响。高尔基体运动的最佳肌球蛋白抑制剂也能够阻止线粒体的运动。在我们的系统中,虽然观察到的过量胞质信号可能掩盖了与细胞器表面结合的肌球蛋白分子,但未发现这些肌球蛋白与高尔基体之间有特异性共定位。从截短肌球蛋白的增强绿色荧光蛋白融合物存在时发现的保存完好的肌动蛋白丝以及肌球蛋白斑点的运动情况来看,我们得出结论,基于肌动球蛋白的胞质环流并未完全停止。综上所述,我们的数据表明上述肌球蛋白直接或间接参与了植物细胞中高尔基体和线粒体的运动。