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Int. J. Mol. Sci. 2018, 19(2), 569; https://doi.org/10.3390/ijms19020569

Effects of TROL Presequence Mutagenesis on Its Import and Dual Localization in Chloroplasts

Laboratory for Plant Molecular Biology and Biotechnology, Division of Molecular Biology, Ruđer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia
Current address: Biologie der Pflanzenzelle, Botanisches Institut, Christian-Albrechts-Universität zu Kiel, Am Botanischen Garten 1-9, 24118 Kiel, Germany.
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Received: 11 January 2018 / Revised: 5 February 2018 / Accepted: 11 February 2018 / Published: 14 February 2018
(This article belongs to the Special Issue Chloroplast)
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Abstract

Thylakoid rhodanase-like protein (TROL) is involved in the final step of photosynthetic electron transport from ferredoxin to ferredoxin: NADP+ oxidoreductase (FNR). TROL is located in two distinct chloroplast compartments—in the inner envelope of chloroplasts, in its precursor form; and in the thylakoid membranes, in its fully processed form. Its role in the inner envelope, as well as the determinants for its differential localization, have not been resolved yet. In this work we created six N-terminal amino acid substitutions surrounding the predicted processing site in the presequence of TROL in order to obtain a construct whose import is affected or localization limited to a single intrachloroplastic site. By using in vitro transcription and translation and subsequent protein import methods, we found that a single amino acid exchange in the presequence, Ala67 to Ile67 interferes with processing in the stroma and directs the whole pool of in vitro translated TROL to the inner envelope of chloroplasts. This result opens up the possibility of studying the role of TROL in the chloroplast inner envelope as well as possible consequence/s of its absence from the thylakoids. View Full-Text
Keywords: TROL; protein import; chloroplasts; dual localization; ATP; inner envelope membrane; thylakoids TROL; protein import; chloroplasts; dual localization; ATP; inner envelope membrane; thylakoids
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Vojta, L.; Čuletić, A.; Fulgosi, H. Effects of TROL Presequence Mutagenesis on Its Import and Dual Localization in Chloroplasts. Int. J. Mol. Sci. 2018, 19, 569.

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