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Insertion of a “chloroplast-like” regulatory segment responsible for thiol modulation into γ-subunit of F0F1-ATPase of the cyanobacterium Synechocystis 6803 by mutagenesis of atpC

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Abstract

A regulatory sequence in the γ subunit of the F0F1-ATPase complex of higher plant chloroplasts, responsible for so-called thiol modulation, is absent in the corresponding polypeptides of the cyanobacterial complexes analysed so far. We have modified the atpC gene encoding this γ subunit in Synechocystis 6803 by site-directed mutagenesis. A segment was introduced coding for nine additional amino acids, including the two functional cysteines, which constitutes the sequence of the respective element in the chloroplast γ subunit. The growth rate as well as the rate of photosynthesis of the transformant was comparable to that of the wild-type, but the transitory increase in respiration observed immediately after a period of illumination was significantly lower in the mutant than in the wild-type. The F1 subcomplex solubilized from thylakoid membranes of both the wild-type and the transformant can be activated by trypsin to yield Ca2+-dependent ATPase activity, but only the F1 from the transformant can be activated by the thiol reagent dithiothreitol.

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Authors and Affiliations

  1. Institut für Biochemie der Pflanzen, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany

    Sabine Werner-Grüne, Dorothea Gunkel, Jürgen Schumann & Heinrich Strotmann

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  1. Sabine Werner-Grüne
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  2. Dorothea Gunkel
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  3. Jürgen Schumann
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  4. Heinrich Strotmann
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Communicated by R. Herrmann

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Werner-Grüne, S., Gunkel, D., Schumann, J. et al. Insertion of a “chloroplast-like” regulatory segment responsible for thiol modulation into γ-subunit of F0F1-ATPase of the cyanobacterium Synechocystis 6803 by mutagenesis of atpC . Molec. Gen. Genet. 244, 144–150 (1994). https://doi.org/10.1007/BF00283515

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  • DOI: https://doi.org/10.1007/BF00283515

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