Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
复制标题
DOI：
10.7554/elife.44558
复制
发表时间：
2019-04-05
期刊：
影响因子：
7.7
通讯作者：
Gendron, Joshua M.
中科院分区：
文献类型：
--
作者：
Feke, Ann;Liu, Wei;Gendron, Joshua M.
生物钟依赖于生物钟蛋白的调节降解来维持节律性。尽管如此，我们知道的介导蛋白质降解的成分很少。这是由于植物E3泛素连接酶家族中高水平的功能冗余。为了克服这个问题，并发现E3泛素连接酶，控制昼夜节律功能，我们产生了一个库的转基因拟南芥植物表达显性负'诱饵' E3泛素连接酶。我们确定了它们对生物钟的影响，并确定了数十种新的生物钟功能的潜在调节因子。为了证明诱饵筛选方法克服冗余并识别真正的时钟调节器的效力，我们对MAC3A（PUB59）和MAC3B（PUB60）进行了后续研究。我们表明，他们冗余控制昼夜节律周期调节剪接。这项工作证明了泛素连接酶诱饵作为筛选平台的可行性，以克服遗传挑战并发现调节植物发育的E3泛素连接酶。
The circadian clock relies on regulated degradation of clock proteins to maintain rhythmicity. Despite this, we know few components that mediate protein degradation. This is due to high levels of functional redundancy within plant E3 ubiquitin ligase families. In order to overcome this issue and discover E3 ubiquitin ligases that control circadian function, we generated a library of transgenic Arabidopsis plants expressing dominant-negative 'decoy' E3 ubiquitin ligases. We determined their effects on the circadian clock and identified dozens of new potential regulators of circadian function. To demonstrate the potency of the decoy screening methodology to overcome redundancy and identify bona fide clock regulators, we performed follow-up studies on MAC3A (PUB59) and MAC3B (PUB60). We show that they redundantly control circadian period by regulating splicing. This work demonstrates the viability of ubiquitin ligase decoys as a screening platform to overcome genetic challenges and discover E3 ubiquitin ligases that regulate plant development.