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Localization and function of SulP, a nuclear-encoded chloroplast sulfate permease in Chlamydomonas reinhardtii

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Abstract

Recent work [H.-C. Chen et al. (2003) Planta 218:98–106] reported on the genomic, proteomic, phylogenetic and evolutionary aspects of a putative nuclear gene (SulP) encoding a chloroplast sulfate permease in the model green alga Chlamydomonas reinhardtii. In this article, evidence is provided for the envelope localization of the SulP protein and its function in the uptake and assimilation of sulfate by the chloroplast. Localization of the SulP protein in the chloroplast envelope was concluded upon isolation of C. reinhardtii chloroplasts, followed by fractionation into envelope and thylakoid membranes and Western blotting of these fractions with specific polyclonal antibodies raised against the recombinant SulP protein. The function of the SulP protein was probed in antisense transformants of C. reinhardtii having lower expression levels of the SulP gene. Results showed that cellular sulfate uptake capacity was lowered as a consequence of attenuated SulP gene expression in the cell, directly affecting rates of de novo protein biosynthesis in the chloroplast. The antisense transformants exhibited phenotypes of sulfate-deprived cells, displaying slow rates of light-saturated oxygen evolution, low levels of Rubisco in the chloroplast and low steady-state levels of the photosystem-II D1 reaction-center protein. The role of the chloroplast sulfate transport in the uptake and assimilation of sulfate in C. reinhardtii is discussed along with its impact on the repair of photosystem-II from a frequently occurring photo-oxidative damage and potential use for the elucidation of the H2-evolution-related metabolism in this green alga.

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Abbreviations

Chl :

Chlorophyll

D1 :

Photosystem-II reaction-center protein

LHCII :

Chl a/b light-harvesting complex of PSII

PS :

Photosystem

QRT–PCR :

Quantitative reverse transcription–polymerase chain reaction

RbcL :

Large subunit of Rubisco

RbcS :

Small subunit of Rubisco

Rubisco :

Ribulose 1,5-bisphosphate carboxylase–oxygenase

SulP :

Sulfate permease

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Acknowledgements

The work was supported in part by an USDA-NRI FD-2002-35100-12278-MELI-08/04 grant and in part by a DaimlerChrysler-UC Berkeley R&D contract.

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  1. Department of Plant and Microbial Biology, 111 Koshland Hall, University of California, Berkeley, CA 94720-3102, USA

    Hsu-Ching Chen & Anastasios Melis

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  1. Hsu-Ching Chen
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  2. Anastasios Melis
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Correspondence to Anastasios Melis.

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Chen, HC., Melis, A. Localization and function of SulP, a nuclear-encoded chloroplast sulfate permease in Chlamydomonas reinhardtii. Planta 220, 198–210 (2004). https://doi.org/10.1007/s00425-004-1331-5

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