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An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice
Various chloroplast transit peptides (CTP) have been used to successfully target some foreign proteins into chloroplasts, but for other proteins these same CTPs have reduced localization efficiencies or fail completely. The underlying cause of the failures remains an open question, and more effectiv...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5387683/ https://www.ncbi.nlm.nih.gov/pubmed/28397859 http://dx.doi.org/10.1038/srep46231 |
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author | Shen, Bo-Ran Zhu, Cheng-Hua Yao, Zhen Cui, Li-Li Zhang, Jian-Jun Yang, Cheng-Wei He, Zheng-Hui Peng, Xin-Xiang |
author_facet | Shen, Bo-Ran Zhu, Cheng-Hua Yao, Zhen Cui, Li-Li Zhang, Jian-Jun Yang, Cheng-Wei He, Zheng-Hui Peng, Xin-Xiang |
author_sort | Shen, Bo-Ran |
collection | PubMed |
description | Various chloroplast transit peptides (CTP) have been used to successfully target some foreign proteins into chloroplasts, but for other proteins these same CTPs have reduced localization efficiencies or fail completely. The underlying cause of the failures remains an open question, and more effective CTPs are needed. In this study, we initially observed that two E.coli enzymes, EcTSR and EcGCL, failed to be targeted into rice chloroplasts by the commonly-used rice rbcS transit peptide (rCTP) and were subsequently degraded. Further analyses revealed that the N-terminal unfolded region of cargo proteins is critical for their localization capability, and that a length of about 20 amino acids is required to attain the maximum localization efficiency. We considered that the unfolded region may alleviate the steric hindrance produced by the cargo protein, by functioning as a spacer to which cytosolic translocators can bind. Based on this inference, an optimized CTP, named RC2, was constructed. Analyses showed that RC2 can more effectively target diverse proteins, including EcTSR and EcGCL, into rice chloroplasts. Collectively, our results provide further insight into the mechanism of CTP-mediated chloroplastic localization, and more importantly, RC2 can be widely applied in future chloroplastic metabolic engineering, particularly for crop plants. |
format | Online Article Text |
id | pubmed-5387683 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-53876832017-04-12 An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice Shen, Bo-Ran Zhu, Cheng-Hua Yao, Zhen Cui, Li-Li Zhang, Jian-Jun Yang, Cheng-Wei He, Zheng-Hui Peng, Xin-Xiang Sci Rep Article Various chloroplast transit peptides (CTP) have been used to successfully target some foreign proteins into chloroplasts, but for other proteins these same CTPs have reduced localization efficiencies or fail completely. The underlying cause of the failures remains an open question, and more effective CTPs are needed. In this study, we initially observed that two E.coli enzymes, EcTSR and EcGCL, failed to be targeted into rice chloroplasts by the commonly-used rice rbcS transit peptide (rCTP) and were subsequently degraded. Further analyses revealed that the N-terminal unfolded region of cargo proteins is critical for their localization capability, and that a length of about 20 amino acids is required to attain the maximum localization efficiency. We considered that the unfolded region may alleviate the steric hindrance produced by the cargo protein, by functioning as a spacer to which cytosolic translocators can bind. Based on this inference, an optimized CTP, named RC2, was constructed. Analyses showed that RC2 can more effectively target diverse proteins, including EcTSR and EcGCL, into rice chloroplasts. Collectively, our results provide further insight into the mechanism of CTP-mediated chloroplastic localization, and more importantly, RC2 can be widely applied in future chloroplastic metabolic engineering, particularly for crop plants. Nature Publishing Group 2017-04-11 /pmc/articles/PMC5387683/ /pubmed/28397859 http://dx.doi.org/10.1038/srep46231 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Shen, Bo-Ran Zhu, Cheng-Hua Yao, Zhen Cui, Li-Li Zhang, Jian-Jun Yang, Cheng-Wei He, Zheng-Hui Peng, Xin-Xiang An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title | An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title_full | An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title_fullStr | An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title_full_unstemmed | An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title_short | An optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
title_sort | optimized transit peptide for effective targeting of diverse foreign proteins into chloroplasts in rice |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5387683/ https://www.ncbi.nlm.nih.gov/pubmed/28397859 http://dx.doi.org/10.1038/srep46231 |
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