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Analysis of Protein Import into Chloroplasts Isolated from Stressed Plants
Published on: November 1, 2016
DEFECTIVE CHLOROPLASTS and LEAVES is essential for chloroplast ribosome RNA processing in Arabidopsis thaliana
Fudan Gao1, Junhao Li1, Qiang Xin1
1Shanghai Engineering Research Center of Plant Germplasm Resources, Shanghai Collaborative Innovation Center of Plant Germplasm Resources Development, College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China.
None:
Chloroplast ribosome biogenesis is essential for plastid development, yet the regulatory mechanisms underlying chloroplast rRNA maturation remain incompletely understood. Here, we characterize the role of DEFECTIVE CHLOROPLASTS AND LEAVES (DCL) in chloroplast rRNA processing in Arabidopsis thaliana. DCL is conserved in photosynthetic eukaryotes and localizes to chloroplasts, whereas its paralogs exhibit distinct localization patterns including chloroplast-mitochondrial dual localization and nuclear-specific targeting. Loss of DCL results in seedling lethality and an ivory leaf phenotype, accompanied by severely decreased accumulation of chloroplast ribosomes. RNA blotting and circularized RT-PCR analyses reveal that DCL is required for the accurate processing of the 16S rRNA and the 23S-4.5S dicistronic precursor as well as for the proper 'hidden break' processing of 23S rRNA. Yeast two-hybrid assays demonstrate that DCL physically interacts with three key factors involved in chloroplast rRNA processing including the mini-RNase III-like enzymes RNC3/RNC4, ribonuclease RNase J (RNJ), and endonuclease YbeY. Thus, our results indicate that DCL is a critical eukaryotic-specific factor that coordinates early endonucleolytic cleavage steps during chloroplast rRNA maturation, thereby enabling proper ribosome assembly and chloroplast translation.
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