Time-resolved Interaction Proteomics of the Putative Scaffold Protein GIGANTEA in Arabidopsis thaliana
Time-resolved Interaction Proteomics of the Putative Scaffold Protein GIGANTEA in Arabidopsis thaliana
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DOI：
10.1101/162271
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发表时间：
2017-08
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通讯作者：
J. Krahmer;Greg S Goralogia;Akane Kubota;Richard S. Johnson;Young Hun Song;M. MacCoss;T. LeBihan;K. Halliday;T. Imaizumi;A. Millar
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作者：
J. Krahmer;Greg S Goralogia;Akane Kubota;Richard S. Johnson;Young Hun Song;M. MacCoss;T. LeBihan;K. Halliday;T. Imaizumi;A. Millar
植物特有的大蛋白 GIGANTEA (GI) 参与许多生理过程，部分通过 GI RNA 表达的昼夜节律和光调节来介导节律性翻译后调节。 GI 结合多种与生物钟、光周期开花控制和非生物胁迫反应有关的蛋白质，并具有共伴侣活性。推而广之，进一步的相互作用伙伴可能会介导不太为人所知的 GI 作用，但这些相互作用者的数量和节律性尚不清楚。在这里，我们利用转基因拟南芥植物组成型表达表位标记的 GI 蛋白的时间序列研究中的定量蛋白质组学，以特定时间的方式寻找潜在的相互作用因子。检测到先前鉴定的 GI 直接和间接相互作用因子，但没有检测到与已知 GI 伙伴 ZTL/FKF1/LKP2 相关的其他 F-box 蛋白。主要的非节律性相互作用蛋白与蛋白质折叠或降解、代谢和染色质修饰有关，包括与其他时钟相关蛋白共享的一小部分伙伴。我们将转录因子同源物命名为 CYCLING DOF FACTOR 6 (CDF6)，它与 GI 和 ZTL/FKF1/LKP2 蛋白相互作用并控制光周期开花。我们的结果表明，除了昼夜节律和开花调节之外，生化途径可能会受到 GIGANTEA 的节律性翻译后控制的影响。意义陈述 最多两句话的意义陈述，总字数不超过 75 个；拟南芥的 GIGANTEA 蛋白已知具有昼夜节律和开花功能，由胃肠道相互作用 F-box 蛋白的 FKF1/LKP2/ZTL 家族介导，然后具有未知范围的共伴侣活性。我们进行了时间分辨相互作用蛋白质组学，将 CDF6 (At1g26790) 识别为控制开花时间的早晨特异性 GI 相互作用因子。与 FKF1 和 CDF 蛋白不同，240 个候选蛋白中的大多数都没有节律性富集。他们将胃肠道与可能介导胃肠道生理功能的蛋白质稳态和代谢功能联系起来。
The large, plant-specific protein GIGANTEA (GI) is involved in many physiological processes, mediating rhythmic, post-translational regulation in part through circadian and light regulation of GI RNA expression. GI binds several proteins implicated in the circadian clock, the control of photoperiodic flowering, and abiotic stress responses, and has co-chaperone activity. By extension, further interaction partners might mediate the less well-understood roles of GI but the number and rhythmicity of these interactors is unknown. Here, we seek potential interactors in a time-specific manner, using quantitative proteomics from a time series study of transgenic Arabidopsis thaliana plants constitutively expressing an epitope-tagged GI protein. Previously-identified, direct and indirect interactors of GI were detected but no further F-box proteins related to known GI partners ZTL/FKF1/LKP2. The predominantly non-rhythmic, interacting proteins were implicated in protein folding or degradation, metabolism and chromatin modification, including a small set of partners shared with other clock-related proteins. A transcription factor homologue that we name CYCLING DOF FACTOR 6 (CDF6) was shown to interact both with GI and the ZTL/FKF1/LKP2 proteins and to control photoperiodic flowering. Our results indicate the biochemical pathways, beyond circadian and flowering regulation, that might be affected by GIGANTEA’s rhythmic, post-translational control. Significance Statement Significance statement of up to two sentences of no more than 75 words total; The GIGANTEA protein of Arabidopsis was known for circadian and flowering functions, mediated by the FKF1/LKP2/ZTL family of GI-interacting, F-box proteins, then for a co-chaperone activity of unknown scope. We performed time-resolved, interaction proteomics, identifying CDF6 (At1g26790) as a morning-specific GI interactor that controls flowering time. Unlike FKF1 and CDF proteins, most of the 240 candidate partners were not rhythmically enriched. They link GI to proteostasis and metabolic functions that might mediate GI’s physiological functions.