1. The checkpoint kinase <scp>TOR</scp> (target of rapamycin) regulates expression of a nuclear‐encoded chloroplast RelA‐SpoT homolog ( <scp>RSH</scp> ) and modulates chloroplast ribosomal <scp>RNA</scp> synthesis in a unicellular red alga
- Author
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Tokiaki Takemura, Yuhta Nomura, Imran Pancha, Keiko Taki, Kazuki Toguchi, Yuzuru Tozawa, Kan Tanaka, and Sousuke Imamura
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0301 basic medicine ,biology ,food and beverages ,RNA ,Cell Biology ,Plant Science ,Ribosomal RNA ,biology.organism_classification ,Ribosome ,RRNA transcription ,Cell biology ,Chloroplast ,Chloroplast ribosome ,03 medical and health sciences ,030104 developmental biology ,Cyanidioschyzon merolae ,Transcription (biology) ,Genetics - Abstract
Chloroplasts are plant organelles that carry out oxygenic photosynthesis. Chloroplast biogenesis depends upon chloroplast ribosomes and their translational activity. However, regulation of chloroplast ribosome biogenesis remains an important unanswered question. In this study, we found that inhibition of target of rapamycin (TOR), a general eukaryotic checkpoint kinase, results in a decline in chloroplast ribosomal RNA (rRNA) transcription in the unicellular red alga, Cyanidioschyzon merolae. Upon TOR inhibition, transcriptomics and other analyses revealed increased expression of a nuclear-encoded chloroplast RelA-SpoT homolog (RSH) gene (CmRSH4b), which encodes a homolog of the guanosine 3'-diphosphate 5'-diphosphate (ppGpp) synthetases that modulate rRNA synthesis in bacteria. Using an Escherichia coli mutant lacking ppGpp, CmRSH4b was demonstrated to have ppGpp synthetase activity. Expression analysis of a green fluorescent protein-fused protein indicated that CmRSH4b localizes to the chloroplast, and overexpression of the CmRSH4b gene resulted in a decrease of chloroplast rRNA synthesis concomitant with growth inhibition and reduction of chloroplast size. Biochemical analyses using C. merolae cell lysates or purified recombinant proteins revealed that ppGpp inhibits bacteria-type RNA polymerase-dependent chloroplast rRNA synthesis as well as a chloroplast guanylate kinase. These results suggest that CmRSH4b-dependent ppGpp synthesis in chloroplasts is an important regulator of chloroplast rRNA transcription. Nuclear and mitochondrial rRNA transcription were both reduced by TOR inhibition, suggesting that the biogeneses of the three independent ribosome systems are interconnected by TOR in plant cells.
- Published
- 2018
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