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AtObgC, a plant ortholog of bacterial Obg, is a chloroplast-targeting GTPase essential for early embryogenesis
Authors:Woo Young Bang  Akira Hata  In Sil Jeong  Tetsuya Umeda  Takayuki Masuda  Ji Chen  Ishizaki Yoko  I Nengah Suwastika  Dae Won Kim  Chak Han Im  Byung Hyun Lee  Yuno Lee  Keun Woo Lee  Takashi Shiina  Jeong Dong Bahk
Institution:1. Division of Applied Life Sciences (BK21 and EB-NCRC), Graduate School of Gyeongsang National University, Jinju, 660-701, Korea
2. Graduate School of Human and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Sakyo-ku, Kyoto, 606-8522, Japan
3. Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan
Abstract:Obg is a ribosome-associated GTPase essential for bacterial viability and is conserved in most organisms, from bacteria to eukaryotes. Obg is also expressed in plants, which predicts an important role for this molecule in plant viability; however, the functions of the plant Obg homologs have not been reported. Here, we first identified Arabidopsis AtObgC as a plant chloroplast-targeting Obg and elucidated its molecular biological and physiological properties. AtObgC encodes a plant-specific Obg GTPase that contains an N-terminal region for chloroplast targeting and has intrinsic GTP hydrolysis activity. A targeting assay using a few AtObgC N-terminal truncation mutants revealed that AtObgC localizes to chloroplasts and its transit peptide consists of more than 50 amino acid residues. Interestingly, GFP-fused full-length AtObgC exhibited a punctate staining pattern in chloroplasts of Arabidopsis protoplasts, which suggests a dimerization or multimerization of AtObgC. Moreover, its Obg fold was indispensable for the generation of the punctate staining pattern, and thus, was supposed to be important for such oligomerization of AtObgC by mediating the protein–protein interaction. In addition, the T-DNA insertion AtObgC null mutant exhibited an embryonic lethal phenotype that disturbed the early stage of embryogenesis. Altogether, our results provide a significant implication that AtObgC as a chloroplast targeting GTPase plays an important role at the early embryogenesis by exerting its function in chloroplast protein synthesis.
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