Inoue Shin-ichiro, Kinoshita Toshinori, Shimazaki Ken-ichiro
Department of Biology, Faculty of Science, Kyushu University, Ropponmatsu, Fukuoka 810-8560, Japan.
Plant Physiol. 2005 Aug;138(4):1994-2004. doi: 10.1104/pp.105.062026. Epub 2005 Jul 22.
The leaf of kidney bean (Phaseolus vulgaris) moves in response to blue light. The movement is induced by a decrease in the turgor pressure of pulvinar motor cells on the irradiated side. In this study, we investigated the initial event of the movement with respect to function of phototropin and the plasma membrane H+-ATPase in the motor cells. The results indicated that, in dark conditions, phototropin existed in a dephosphorylated state and the H+-ATPase existed in a phosphorylated state. A pulse of blue light (30 s) induced the phosphorylation of phototropin and the dephosphorylation of the H+-ATPase as determined by the binding behavior of 14-3-3 protein. Phototropin phosphorylation occurred rapidly, followed by the transient gradual dephosphorylation of the H+-ATPase. When the specific flavoprotein inhibitor diphenyleneiodonium and the protein kinase inhibitors K-252a and staurosporine were administered to pulvinar cells, both phototropin phosphorylation and H+-ATPase dephosphorylation were inhibited. The phosphorylation and dephosphorylation exhibited similar fluence rate dependencies to blue light. These results indicated that phototropin may function upstream of the plasma membrane H+-ATPase and decrease the activity of H+-ATPase by dephosphorylation. We provide evidence for the existence of three kinds of phototropins in pulvinar motor cells.
菜豆(Phaseolus vulgaris)的叶片会对蓝光做出反应。这种运动是由受照射一侧叶枕运动细胞膨压的降低所诱导的。在本研究中,我们针对叶枕运动细胞中向光素和质膜H⁺-ATP酶的功能,研究了这种运动的初始事件。结果表明,在黑暗条件下,向光素以去磷酸化状态存在,而H⁺-ATP酶以磷酸化状态存在。如通过14-3-3蛋白的结合行为所测定的,一次蓝光脉冲(30秒)诱导了向光素的磷酸化和H⁺-ATP酶的去磷酸化。向光素磷酸化迅速发生,随后H⁺-ATP酶出现短暂的逐渐去磷酸化。当向叶枕细胞施用特异性黄素蛋白抑制剂二苯碘鎓以及蛋白激酶抑制剂K-252a和星形孢菌素时,向光素磷酸化和H⁺-ATP酶去磷酸化均受到抑制。磷酸化和去磷酸化对蓝光表现出相似的光通量率依赖性。这些结果表明,向光素可能在质膜H⁺-ATP酶的上游发挥作用,并通过去磷酸化降低H⁺-ATP酶的活性。我们提供了叶枕运动细胞中存在三种向光素的证据。