TOR kinase activity is decreased by sugar depletion in liquid culture system
In mammals and yeasts, the target of rapamycin (TOR) is a central regulator in glucose signaling to promote cell proliferation and growth. However, in plant, the investigation of sugar (glucose) -TOR signaling has been hindered by the innate capacity of autotrophic plants to produce carbon-related nutrients via photosynthesis. Here, we established a protocol which provided a manipulated liquid culture system for efficient depletion of the endogenous carbon nutrients in Arabidopsis seedlings. Exogenously sugar (glucose) could then be supplied to investigate the role of sugar (glucose)-TOR signaling in cell division and growth.
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To investigate the effect of sugar starvation on endogenous TOR-kinase activity, we monitored TOR kinase activity based on the phosphorylation of T449 in the TOR-conserved substrate ribosomal S6 kinase 1 (S6K1) by common Western blot method.