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Using Microtiter Dish Radiolabeling for Multiple In Vivo Measurements Of Escherichia coli pppGpp Followed by Thin Layer Chromatography
Published on: June 4, 2019
Loss of the plastidial signaling molecule, ppGpp, accumulation alters nuclear gene expression during nitrogen
Takanari Nemoto1,2, Yuto Omata1, Masataka Inazu1
1Department of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
Abstract:
Nitrogen (N) deficiency triggers major transcriptional reprogramming in plants. The chloroplast alarmone, guanosine tetraphosphate (ppGpp) synthesized and hydrolyzed by RelA-SpoT homologs (RSHs), has been proposed to regulate cellular metabolism. Here, we characterized an Arabidopsis mutant lacking all RSHs (quadruple), which accumulates no detectable ppGpp. Transcriptome analysis showed that 774 and 2,928 nuclear-encoded genes were differentially expressed in quadruple compared with the wild type (WT), under +N and -N conditions, respectively. Upon transition from +N to -N conditions, 2,487 nuclear genes in WT and 1,505 in quadruple primarily associated with cell wall biosynthesis and defense responses, were differentially expressed, suggesting that plastidial ppGpp is involved in the reprogramming of nuclear gene expression in response to N availability. Network analysis of transcription factors (TFs) indicated that ppGpp alters TFs expression and identified 11 candidates of master regulator of TFs expression by ppGpp-dependent manner during N starvation. Transcript levels of several plastid-encoded genes were higher in quadruple compared to WT under -N conditions, whereas mitochondrial transcripts were less affected. Together, these findings suggest that ppGpp acts as both a regulator and a key component of retrograde signaling, coordinating nuclear transcription and metabolic adaptation in response to N availability.
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