A proteomic screen reveals sCFGrr1 targets that regulate the glycolytic-gluconeogenic switch
A proteomic screen reveals sCFGrr1 targets that regulate the glycolytic-gluconeogenic switch
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DOI：
10.1038/ncb1639
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发表时间：
2007-10-01
影响因子：
21.3
通讯作者：
Toczyski, David P.
中科院分区：
文献类型：
--
作者：
Benanti, Jennifer A.;Cheung, Stephanie K.;Toczyski, David P.
进入细胞周期是由营养物质的可用性调节的，这样当资源有限时细胞就不会分裂。Skp 1-Cul 1-F-box（SCF）泛素连接酶与F-box蛋白Grr 1（SCFGrr 1）控制细胞周期进入和葡萄糖传感器的调节剂的蛋白水解营业额，这表明它将细胞周期与营养物质的可用性联系起来。在这里，我们表明，SCFGrr 1广泛调节细胞代谢。我们开发了一种蛋白质组学筛选方法，该方法使用高通量定量显微镜来全面筛选泛素连接酶底物。SCFGrr 1的7个新的代谢靶点被确定，包括糖酵解的两个调节因子-转录因子Tye 7和Pfk 27。后者产生第二信使果糖-2，6-二磷酸，其激活糖酵解并抑制糖异生。我们表明，SCFGrr 1的目标Pfk 27和Tye 7响应葡萄糖去除。此外，Pfk 27被激酶Snf 1磷酸化，并且不可磷酸化的Pfk 27在不存在葡萄糖的情况下是稳定的并且抑制生长。这些结果证明了SCFGrr 1在调节糖酵解-促代谢开关中的作用。
Entry into the cell cycle is regulated by nutrient availability such that cells do not divide when resources are limited. The Skp1-Cul1-F-box(SCF) ubiquitin ligase with the F-box protein Grr1 ( SCFGrr1) controls the proteolytic turnover of regulators of cell-cycle entry and a glucose sensor, suggesting that it links the cell cycle with nutrient availability. Here, we show that SCFGrr1 broadly regulates cellular metabolism. We have developed a proteomic screening method that uses high-throughput quantitative microscopy to comprehensively screen for ubiquitin-ligase substrates. Seven new metabolic targets of SCFGrr1 were identified, including two regulators of glycolysis-the transcription factor Tye7 and Pfk27. The latter produces the second messenger fructose-2,6- bisphosphate that activates glycolysis and inhibits gluconeogenesis. We show that SCFGrr1 targets Pfk27 and Tye7 in response to glucose removal. Moreover, Pfk27 is phosphorylated by the kinase Snf1, and unphosphorylatable Pfk27 is stable and inhibits growth in the absence of glucose. These results demonstrate a role for SCFGrr1 in regulating the glycolytic-gluconeogenic switch.