COLD REGULATED GENE 27 and 28 antagonize the transcriptional activity of the RVE8/LNK1/LNK2 circadian complex.
COLD REGULATED GENE 27 and 28 antagonize the transcriptional activity of the RVE8/LNK1/LNK2 circadian complex.
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DOI：
10.1093/plphys/kiad210
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发表时间：
2023-07-03
期刊：
影响因子：
7.4
通讯作者：
Nusinow, Dmitri A.
中科院分区：
文献类型：
--
作者：
Sorkin, Maria L.;Tzeng, Shin-Cheng;King, Stefanie;Romanowski, Andres;Kahle, Nikolai;Bindbeutel, Rebecca;Hiltbrunner, Andreas;Yanovsky, Marcelo J.;Evans, Bradley S.;Nusinow, Dmitri A.
植物中的许多分子和生理过程发生在一天中的特定时间。这些每日节律部分由昼夜节律钟协调，昼夜节律钟是一种计时器，它利用白昼长度和温度来维持各种时钟调节表型中的24小时节律。昼夜节律MYB样转录因子REVEILLE 8（RVE 8）与其转录辅激活因子NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 1（LNK 1）和LNK 2相互作用以促进晚相时钟基因和耐寒因子的表达。虽然遗传学方法通常用于发现时钟内以及时钟元件与其他途径之间的联系，但在这里，我们使用亲和纯化与质谱联用（APMS）来鉴定拟南芥（Arabidopsis thaliana）中RVE 8-LNK 1/LNK 2复合物的时间特异性蛋白质相互作用物。在RVE 8/LNK 1/LNK 2的相互作用物中，冷调节基因27（COR 27）和COR 28以晚上特异性方式共沉淀。除了COR 27和COR 28之外，我们还发现了与温度相关的相互作用物的富集，这使我们建立了LNK 1和LNK 2在时钟温度夹带中的先前未表征的作用。我们确定RVE 8、LNK 1和COR 27或COR 28在酵母（酿酒酵母）中形成三方复合物，并且这种相互作用在植物中的作用用于拮抗RVE 8靶基因的转录激活，可能通过介导RVE 8蛋白在晚上降解。总之，这些结果说明了蛋白质组学方法可以用来确定一天中特定时间的蛋白质相互作用。RVE 8-LNK-COR蛋白复合物的发现表明了一种以前未知的昼夜节律和温度信号通路的调节机制。冷调节基因27和28结合并拮抗昼夜节律钟相关的RVE 8/LNK复合物在拟南芥中诱导靶基因在晚上表达的能力。
Many molecular and physiological processes in plants occur at a specific time of day. These daily rhythms are coordinated in part by the circadian clock, a timekeeper that uses daylength and temperature to maintain rhythms of ∼24 h in various clock-regulated phenotypes. The circadian MYB-like transcription factor REVEILLE 8 (RVE8) interacts with its transcriptional coactivators NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 1 (LNK1) and LNK2 to promote the expression of evening-phased clock genes and cold tolerance factors. While genetic approaches have commonly been used to discover connections within the clock and between clock elements and other pathways, here, we used affinity purification coupled with mass spectrometry (APMS) to identify time-of-day–specific protein interactors of the RVE8-LNK1/LNK2 complex in Arabidopsis (Arabidopsis thaliana). Among the interactors of RVE8/LNK1/LNK2 were COLD-REGULATED GENE 27 (COR27) and COR28, which coprecipitated in an evening-specific manner. In addition to COR27 and COR28, we found an enrichment of temperature-related interactors that led us to establish a previously uncharacterized role for LNK1 and LNK2 in temperature entrainment of the clock. We established that RVE8, LNK1, and either COR27 or COR28 form a tripartite complex in yeast (Saccharomyces cerevisiae) and that the effect of this interaction in planta serves to antagonize transcriptional activation of RVE8 target genes, potentially through mediating RVE8 protein degradation in the evening. Together, these results illustrate how a proteomic approach can be used to identify time-of-day–specific protein interactions. Discovery of the RVE8-LNK-COR protein complex indicates a previously unknown regulatory mechanism for circadian and temperature signaling pathways. COLD REGULATED GENE 27 and 28 bind to and antagonize the ability of the circadian clock–associated RVE8/LNK complex to induce target gene expression in the evening in Arabidopsis thaliana.
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