Collaborative Research: An F-box protein targeting PIF1 and PIF3

光是影响植物发育最重要的环境因子之一，除了驱动光合作用外，还控制着种子萌发、生长、绿化等许多植物发育过程。植物科学的一个核心问题是，在细胞水平上，植物是如何对光作出反应的？已知某些蛋白质充当光受体，而其他蛋白质充当光反应的抑制剂。在黑暗中，这些蛋白质在细胞内保持分离，但在光照下，它们到达同一个目的地——细胞核。受体的接近以某种方式通过第三种先前未识别的中介蛋白的作用破坏了抑制剂。本研究旨在澄清介质的身份，并研究它如何影响抑制剂的稳定性。预计从植物细胞中分离出抑制蛋白，使其拥有比通常更多的抑制蛋白，并暴露在光线下，将同时分离出介质蛋白，因为它们彼此具有亲和力，反之亦然。这些在体内的蛋白质-蛋白质相互作用将被证明使用抗体针对这两种蛋白质。这种相互作用也将使用荧光标记的抑制剂和中介蛋白来可视化，只有当抑制剂和中介蛋白处于物理接触时才可见。只有在植物细胞暴露于光下才会产生可见信号，并且随着抑制剂的破坏，该信号会随着时间的推移而减弱。博士后合作者将获得该奖项的支持，因为他们率先阐明了植物对光反应的这一方面。在细胞水平上对光感知的更深入的了解，将为设计作物植物提供机会，使其更快发芽，在田间建立更大的林分，并增强植物高度和结构的改变。

Light is one of the most important environmental factors influencing plant development, controlling seed germination, growth, greening, and many other plant developmental processes in addition to driving photosynthesis. A central question in plant science concerns how, at the cellular level, the plant responds to light? Certain proteins are known to act as light receptors while others act as inhibitors of light responses. In darkness these proteins are kept separate within the cell but, upon illumination, arrive at the same destination, the nucleus. The receptor proximity somehow destroys the inhibitor through the action of a third, previously unidentified, mediator protein. This research seeks to clarify the identity of the mediator and investigate how it influences the stability of the inhibitor. It is anticipated that isolating the inhibitor protein from plant cells engineered to possess greater than usual amounts of the inhibitor protein, and exposed to light, will simultaneously isolate the mediator protein due to their affinity for one another while the converse will also be true. These in vivo protein-protein interactions will be demonstrated using antibodies against both proteins. This interaction will also be visualized using fluorescently tagged inhibitor and mediator proteins that are visible only when the inhibitor and mediator proteins are in physical contact. Visible signal should be generated only following exposure of the plant cells to light and this signal should then attenuate over time as the inhibitor is destroyed. A post-doctoral collaborator will be supported by this award as they spearhead efforts to elucidate this facet of the plant response to light. A greater understanding of light perception at the cellular level should provide opportunities to engineer crop plants for faster germination, greater stand establishment in the field, and potentiate the alteration of plant stature and structure.