Blue-light-activated phototropin2 trafficking from the cytoplasm to Golgi/post-Golgi vesicles.
Blue-light-activated phototropin2 trafficking from the cytoplasm to Golgi/post-Golgi vesicles.
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DOI：
10.1093/jxb/eru172
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发表时间：
2014-07
影响因子：
6.9
通讯作者：
Gabryś H
中科院分区：
文献类型：
--
作者：
Aggarwal C;Banaś AK;Kasprowicz-Maluśki A;Borghetti C;Labuz J;Dobrucki J;Gabryś H
显示了UVA/蓝光受体光向质2的蓝光诱导的运输。证据提供了两种途径的存在下，一个直接phototropin 2的高尔基体和后高尔基体囊泡，和其他的降解。向光蛋白是质膜定位的UVA/蓝光光受体，其介导向光性、抑制初级下胚轴伸长、叶片定位、叶绿体运动和气孔开放。蓝光照射激活趋光蛋白的C-末端丝氨酸/苏氨酸激酶结构域，其使受体自磷酸化。拟南芥编码两种向光蛋白，phot 1和phot 2。响应蓝光，phot 1从质膜移动到胞质溶胶和phot 2易位到高尔基复合体。在这项研究中，蓝光诱导的phot 2运输的分子机制和途径进行了论证。它表明，Atphot 2表现在一个类似的方式下瞬时表达35 S或其天然启动子。phot 2激酶结构域而不是蓝光介导的自磷酸化是受体易位所必需的。使用共定位和蛋白质印迹，受体被证明从细胞质移动到高尔基复合体，然后到后高尔基结构。该结果得到了布雷菲德菌素A（分泌途径的抑制剂）的证实，它破坏了phot 2的运输。通过双分子荧光互补，观察到phot 2和网格蛋白轻链2之间的关联。在质膜上观察到荧光。使用免疫共沉淀法证实了结果。然而，tyrphostin 23（网格蛋白介导的内吞作用的抑制剂）和渥曼青霉素（受体内吞作用的抑制剂）不能阻止phot 2运输，表明没有参与受体内吞作用的phot 2点状结构的形成。蛋白质周转研究表明，受体在黑暗和蓝光下持续降解。phot 2的降解进行通过转运途径不同的高尔基复合体。
The blue-light-induced trafficking of the UVA/blue light receptor phototropin2 is shown. Evidence is provided for the presence of two pathways, one directing phototropin2 to the Golgi and post-Golgi vesicles, and the other to degradation. Phototropins are plasma membrane-localized UVA/blue light photoreceptors which mediate phototropism, inhibition of primary hypocotyl elongation, leaf positioning, chloroplast movements, and stomatal opening. Blue light irradiation activates the C-terminal serine/threonine kinase domain of phototropin which autophosphorylates the receptor. Arabidopsis thaliana encodes two phototropins, phot1 and phot2. In response to blue light, phot1 moves from the plasma membrane into the cytosol and phot2 translocates to the Golgi complex. In this study the molecular mechanism and route of blue-light-induced phot2 trafficking are demonstrated. It is shown that Atphot2 behaves in a similar manner when expressed transiently under 35S or its native promoter. The phot2 kinase domain but not blue-light-mediated autophosphorylation is required for the receptor translocation. Using co-localization and western blotting, the receptor was shown to move from the cytoplasm to the Golgi complex, and then to the post-Golgi structures. The results were confirmed by brefeldin A (an inhibitor of the secretory pathway) which disrupted phot2 trafficking. An association was observed between phot2 and the light chain2 of clathrin via bimolecular fluorescence complementation. The fluorescence was observed at the plasma membrane. The results were confirmed using co-immunoprecipitation. However, tyrphostin23 (an inhibitor of clathrin-mediated endocytosis) and wortmannin (a suppressor of receptor endocytosis) were not able to block phot2 trafficking, indicating no involvement of receptor endocytosis in the formation of phot2 punctuate structures. Protein turnover studies indicated that the receptor was continuously degraded in both darkness and blue light. The degradation of phot2 proceeded via a transport route different from translocation to the Golgi complex.
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影响因子：
6.9
作者：
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通讯作者：
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DOI：
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