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The chloroplast NifS-like protein of Arabidopsis thaliana is required for iron–sulfur cluster formation in ferredoxin

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Abstract

Plastids are known to be able to synthesize their own iron–sulfur clusters, but the biochemical machinery responsible for this process is not known. In this study it is investigated whether CpNifS, the chloroplastic NifS-like cysteine desulfurase of Arabidopsis thaliana (L.) Heynh. is responsible for the release of sulfur from cysteine for the biogenesis of iron–sulfur (Fe–S) clusters in chloroplasts. Using an in vitro reconstitution assay it was found that purified CpNifS was sufficient for Fe–S cluster formation in ferredoxin in the presence of cysteine and a ferrous iron salt. Antibody-depletion experiments using stromal extract showed that CpNifS is also essential for the Fe–S cluster formation activity of chloroplast stroma. The activity of CpNifS in the stroma was 50- to 80-fold higher than that of purified CpNifS on a per-protein basis, indicating that other stromal factors cooperate in Fe–S cluster formation. When stromal extract was separated on a gel-filtration column, most of the CpNifS eluted as a dimer of 86 kDa, but a minor fraction of the stromal CpNifS eluted at a molecular weight of approx. 600 kDa, suggesting the presence of a multi-protein complex. The possible nature of the interacting proteins is discussed.

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Abbreviations

CpNifS :

Chloroplast NifS-like protein

fd :

Ferredoxin

PLP :

Pyridoxal-5′-phosphate

DTT :

Dithiothreitol

Fe–S :

Iron–sulfur

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Acknowledgement

This work was supported by USDA–NRI Grant # 2003–35318–13758 to E.A.H.P.S. and M.P.

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Authors and Affiliations

  1. Department of Biology, Colorado State University, Anatomy/Zoology Building, Fort Collins, CO 80523, USA

    Hong Ye, Gulnara F. Garifullina, Salah E. Abdel-Ghany, Lihong Zhang, Elizabeth A. H. Pilon-Smits & Marinus Pilon

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  1. Hong Ye
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  2. Gulnara F. Garifullina
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  3. Salah E. Abdel-Ghany
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  4. Lihong Zhang
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  5. Elizabeth A. H. Pilon-Smits
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  6. Marinus Pilon
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Correspondence to Marinus Pilon.

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Ye, H., Garifullina, G.F., Abdel-Ghany, S.E. et al. The chloroplast NifS-like protein of Arabidopsis thaliana is required for iron–sulfur cluster formation in ferredoxin. Planta 220, 602–608 (2005). https://doi.org/10.1007/s00425-004-1388-1

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  • DOI: https://doi.org/10.1007/s00425-004-1388-1

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