Abstract
Aminoacyl-tRNA synthetases (AARSs) play a critical role in translation and are thus required in three plant protein-synthesizing compartments: cytosol, mitochondria and plastids. A systematic study had previously shown extensive sharing of organellar AARSs from Arabidopsis thaliana, mostly between mitochondria and chloroplasts. However, distribution of AARSs from monocot species, such as maize, has never been experimentally investigated. Here we demonstrate dual targeting of maize seryl-tRNA synthetase, SerZMo, into both mitochondria and chloroplasts using combination of complementary methods, including in vitro import assay, transient expression analysis of green fluorescent protein (GFP) fusions and immunodetection. We also show that SerZMo dual localization is established by the virtue of an ambiguous targeting peptide. Full-length SerZMo protein fused to GFP is targeted to chloroplast stromules, indicating that SerZMo protein performs its function in plastid stroma. The deletion mutant lacking N-terminal region of the ambiguous SerZMo targeting peptide was neither targeted into mitochondria nor chloroplasts, indicating the importance of this region in both mitochondrial and chloroplastic import.
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Abbreviations
- AARS:
-
Aminoacyl-tRNA synthetase
- SerRS:
-
Seryl-tRNA synthetase
- GFP:
-
Green fluorescent protein
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Acknowledgments
We thank Claire Pujol for initial help with in vitro import studies, Dr Igor Weber for inital help with confocal microscopy and Dr Hrvoje Fulgosi for the gift of pTH-2 plasmid. J.R.P thanks Dr Laurence Marechal-Drouard for hospitality. Visit of J.R.P to Strasbourg was supported by short-term scholarship of the French government and Croatian Ministry of Science, Education and Sports. This work was supported by grant from the Ministry of Science, Education and Sports of the Republic of Croatia (119-0982913-1358).
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Communicated by J.R. Liu.
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Rokov-Plavec, J., Dulic, M., Duchêne, AM. et al. Dual targeting of organellar seryl-tRNA synthetase to maize mitochondria and chloroplasts. Plant Cell Rep 27, 1157–1168 (2008). https://doi.org/10.1007/s00299-008-0542-9
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DOI: https://doi.org/10.1007/s00299-008-0542-9