State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
Plant Cell. 2011 Mar;23(3):1093-106. doi: 10.1105/tpc.110.082420. Epub 2011 Mar 15.
The involvement of cytoskeleton-related proteins in regulating mitochondrial respiration has been revealed in mammalian cells. However, it is unclear if there is a relationship between the microtubule-based motor protein kinesin and mitochondrial respiration. In this research, we demonstrate that a plant-specific kinesin, Kinesin-like protein 1 (KP1; At KIN14 h), is involved in respiratory regulation during seed germination at a low temperature. Using in vitro biochemical methods and in vivo transgenic cell observations, we demonstrate that KP1 is able to localize to mitochondria via its tail domain (C terminus) and specifically interacts with a mitochondrial outer membrane protein, voltage-dependent anion channel 3 (VDAC3). Targeting of the KP1-tail to mitochondria is dependent on the presence of VDAC3. When grown at 4° C, KP1 dominant-negative mutants (TAILOEs) and vdac3 mutants exhibited a higher seed germination frequency. All germinating seeds of the kp1 and vdac3 mutants had increased oxygen consumption; the respiration balance between the cytochrome pathway and the alternative oxidase pathway was disrupted, and the ATP level was reduced. We conclude that the plant-specific kinesin, KP1, specifically interacts with VDAC3 on the mitochondrial outer membrane and that both KP1 and VDAC3 regulate aerobic respiration during seed germination at low temperature.
细胞骨架相关蛋白在调节哺乳动物细胞中线粒体呼吸中的作用已经被揭示。然而,微管驱动蛋白 kinesin 是否与线粒体呼吸之间存在关系尚不清楚。在这项研究中,我们证明了一种植物特异性的驱动蛋白 Kinesin-like protein 1 (KP1; At KIN14 h) 参与低温下种子萌发过程中的呼吸调节。通过体外生化方法和体内转基因细胞观察,我们证明 KP1 能够通过其尾部结构域(C 端)定位到线粒体,并特异性地与一种线粒体外膜蛋白电压依赖性阴离子通道 3 (VDAC3) 相互作用。KP1 尾部靶向线粒体取决于 VDAC3 的存在。在 4°C 下生长时,KP1 显性负突变体(TAILOEs)和 vdac3 突变体表现出更高的种子萌发频率。kp1 和 vdac3 突变体中所有萌发的种子的耗氧量增加;细胞色素途径和交替氧化酶途径之间的呼吸平衡被打破,ATP 水平降低。我们得出结论,植物特异性的驱动蛋白 KP1 特异性地与线粒体外膜上的 VDAC3 相互作用,并且 KP1 和 VDAC3 都调节低温下种子萌发过程中的有氧呼吸。