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Title: Coexpressed subunits of dual genetic origin define a conserved supercomplex mediating essential protein import into chloroplasts

Abstract

In photosynthetic eukaryotes, thousands of proteins are translated in the cytosol and imported into the chloroplast through the concerted action of two translocons—termed TOC and TIC—located in the outer and inner membranes of the chloroplast envelope, respectively. The degree to which the molecular composition of the TOC and TIC complexes is conserved over phylogenetic distances has remained controversial. Here, we combine transcriptomic, biochemical, and genetic tools in the green alga Chlamydomonas ( Chlamydomonas reinhardtii ) to demonstrate that, despite a lack of evident sequence conservation for some of its components, the algal TIC complex mirrors the molecular composition of a TIC complex from Arabidopsis thaliana. The Chlamydomonas TIC complex contains three nuclear-encoded subunits, Tic20, Tic56, and Tic100, and one chloroplast-encoded subunit, Tic214, and interacts with the TOC complex, as well as with several uncharacterized proteins to form a stable supercomplex (TIC-TOC), indicating that protein import across both envelope membranes is mechanistically coupled. Expression of the nuclear and chloroplast genes encoding both known and uncharacterized TIC-TOC components is highly coordinated, suggesting that a mechanism for regulating its biogenesis across compartmental boundaries must exist. Conditional repression of Tic214, the only chloroplast-encoded subunit in the TIC-TOC complex, impairs the import of chloroplast proteinsmore » with essential roles in chloroplast ribosome biogenesis and protein folding and induces a pleiotropic stress response, including several proteins involved in the chloroplast unfolded protein response. These findings underscore the functional importance of the TIC-TOC supercomplex in maintaining chloroplast proteostasis.« less

Authors:
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Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1730999
Grant/Contract Number:  
FC02-02ER63421
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 117 Journal Issue: 51; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English

Citation Formats

Ramundo, Silvia, Asakura, Yukari, Salomé, Patrice A., Strenkert, Daniela, Boone, Morgane, Mackinder, Luke C. M., Takafuji, Kazuaki, Dinc, Emine, Rahire, Michèle, Crèvecoeur, Michèle, Magneschi, Leonardo, Schaad, Olivier, Hippler, Michael, Jonikas, Martin C., Merchant, Sabeeha, Nakai, Masato, Rochaix, Jean-David, and Walter, Peter. Coexpressed subunits of dual genetic origin define a conserved supercomplex mediating essential protein import into chloroplasts. United States: N. p., 2020. Web. https://doi.org/10.1073/pnas.2014294117.
Ramundo, Silvia, Asakura, Yukari, Salomé, Patrice A., Strenkert, Daniela, Boone, Morgane, Mackinder, Luke C. M., Takafuji, Kazuaki, Dinc, Emine, Rahire, Michèle, Crèvecoeur, Michèle, Magneschi, Leonardo, Schaad, Olivier, Hippler, Michael, Jonikas, Martin C., Merchant, Sabeeha, Nakai, Masato, Rochaix, Jean-David, & Walter, Peter. Coexpressed subunits of dual genetic origin define a conserved supercomplex mediating essential protein import into chloroplasts. United States. https://doi.org/10.1073/pnas.2014294117
Ramundo, Silvia, Asakura, Yukari, Salomé, Patrice A., Strenkert, Daniela, Boone, Morgane, Mackinder, Luke C. M., Takafuji, Kazuaki, Dinc, Emine, Rahire, Michèle, Crèvecoeur, Michèle, Magneschi, Leonardo, Schaad, Olivier, Hippler, Michael, Jonikas, Martin C., Merchant, Sabeeha, Nakai, Masato, Rochaix, Jean-David, and Walter, Peter. Thu . "Coexpressed subunits of dual genetic origin define a conserved supercomplex mediating essential protein import into chloroplasts". United States. https://doi.org/10.1073/pnas.2014294117.
@article{osti_1730999,
title = {Coexpressed subunits of dual genetic origin define a conserved supercomplex mediating essential protein import into chloroplasts},
author = {Ramundo, Silvia and Asakura, Yukari and Salomé, Patrice A. and Strenkert, Daniela and Boone, Morgane and Mackinder, Luke C. M. and Takafuji, Kazuaki and Dinc, Emine and Rahire, Michèle and Crèvecoeur, Michèle and Magneschi, Leonardo and Schaad, Olivier and Hippler, Michael and Jonikas, Martin C. and Merchant, Sabeeha and Nakai, Masato and Rochaix, Jean-David and Walter, Peter},
abstractNote = {In photosynthetic eukaryotes, thousands of proteins are translated in the cytosol and imported into the chloroplast through the concerted action of two translocons—termed TOC and TIC—located in the outer and inner membranes of the chloroplast envelope, respectively. The degree to which the molecular composition of the TOC and TIC complexes is conserved over phylogenetic distances has remained controversial. Here, we combine transcriptomic, biochemical, and genetic tools in the green alga Chlamydomonas ( Chlamydomonas reinhardtii ) to demonstrate that, despite a lack of evident sequence conservation for some of its components, the algal TIC complex mirrors the molecular composition of a TIC complex from Arabidopsis thaliana. The Chlamydomonas TIC complex contains three nuclear-encoded subunits, Tic20, Tic56, and Tic100, and one chloroplast-encoded subunit, Tic214, and interacts with the TOC complex, as well as with several uncharacterized proteins to form a stable supercomplex (TIC-TOC), indicating that protein import across both envelope membranes is mechanistically coupled. Expression of the nuclear and chloroplast genes encoding both known and uncharacterized TIC-TOC components is highly coordinated, suggesting that a mechanism for regulating its biogenesis across compartmental boundaries must exist. Conditional repression of Tic214, the only chloroplast-encoded subunit in the TIC-TOC complex, impairs the import of chloroplast proteins with essential roles in chloroplast ribosome biogenesis and protein folding and induces a pleiotropic stress response, including several proteins involved in the chloroplast unfolded protein response. These findings underscore the functional importance of the TIC-TOC supercomplex in maintaining chloroplast proteostasis.},
doi = {10.1073/pnas.2014294117},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 51,
volume = 117,
place = {United States},
year = {2020},
month = {12}
}

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