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bioRxiv · 10.1101/2020.10.28.360057

Cytidine triphosphate synthase two provides nucleotides for chloroplast development and stress responses

Alamdari, K.·Fisher, K. E.·Welsh, D. W.·Rai, S.·Palos, K.·Nelson, A. D.·Woodson, J. D.
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Abstract

O_LIReactive oxygen species (ROS) produced in chloroplasts cause oxidative damage, but also signal to initiate chloroplast quality control pathways, cell death, and gene expression. The mechanisms behind these signals are largely unknown. C_LIO_LIThe Arabidopsis thaliana plastid ferrochelatase two (fc2) mutant produces the ROS singlet oxygen in chloroplasts that activates such signaling pathways. Here we mapped one fc2 suppressor mutation to CYTIDINE TRIPHOSPHATE SYNTHASE TWO (CTPS2), which encodes one of five enzymes in Arabidopsis necessary for de novo cytoplasmic CTP (and dCTP) synthesis. C_LIO_LIThe ctps2 mutation reduces chloroplast transcripts and DNA content without similarly affecting mitochondria. Chloroplast nucleic acid content and singlet oxygen signaling are restored by exogenous feeding of the dCTP precursor deoxycytidine, suggesting ctps2 blocks signaling by limiting nucleotides for chloroplast genome maintenance. C_LIO_LIAn investigation of CTPS orthologs in Brassicaceae showed CTPS2 is a member of an ancient lineage distinct from CTPS3. Complementation studies confirmed this analysis; CTPS3 was unable to compensate for CTPS2 function in providing nucleotides for chloroplast DNA and signaling. C_LIO_LIOur studies link cytoplasmic nucleotide metabolism with chloroplast quality control pathways. Such a connection is achieved by a conserved clade of CTPS enzymes that may have evolved specialized functions in providing nucleotides to specific subcellular compartments. C_LI

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Alamdari, K., Fisher, K. E., Welsh, D. W., Rai, S., Palos, K., Nelson, A. D., Woodson, J. D.. 2020-10-29. Cytidine triphosphate synthase two provides nucleotides for chloroplast development and stress responses. https://doi.org/10.1101/2020.10.28.360057

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