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J. Biol. Chem., Vol. 279, Issue 47, 48620-48629, November 19, 2004

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Accumulation of the D2 Protein Is a Key Regulatory Step for Assembly of the Photosystem II Reaction Center Complex in Synechocystis PCC 6803^*

Josef Komenda, Veronika Reisinger¶, Bernd Christian Müller||, Marika Dobáková, Bernhard Granvogl¶, and Lutz Andreas Eichacker¶**

From the Institute of Microbiology, Opatovick mln, 379 81 Tebo, Czech Republic, the Institute of Physical Biology, University of South Bohemia, Zámek 136, 370 05 Nové Hrady, Czech Republic, the ^¶Botanisches Institute der Ludwig-Maximilians Universität München, Menzinger Strasse 67, 80368 München, Germany, and ^||Hoffmann-La Roche, RCMG, CH-4070 Basel, Switzerland

Accumulation of monomer and dimer photosystem (PS) II reaction center core complexes has been analyzed by two-dimensional Blue-native/SDS-PAGE in Synechocystis PCC 6803 wild type and in mutant strains lacking genes psbA, psbB, psbC, psbDIC/DII, or the psbEFLJ operon. In vivo pulse-chase radiolabeling experiments revealed that mutant cells assembled PSII precomplexes only. In psbC and psbB, assembly of reaction center cores lacking CP43 and reaction center complexes was detected, respectively. In psbA, protein subunits CP43, CP47, D2, and cytochrome b₅₅₉ were synthesized, but proteins did not assemble. Similarly, in psbD/C lacking D2, and CP43, the de novo synthesized proteins D1, CP47, and cytochrome b₅₅₉ did not form any mutual complexes, indicating that assembly of the reaction center complex is a prerequisite for assembly with core subunits CP47 and CP43. Finally, although CP43 and CP47 accumulated in psbEFLJ, D2 was neither expressed nor accumulated. We, furthermore, show that the amount of D2 is high in the strain lacking D1, whereas the amount of D1 is low in the strain lacking D2. We conclude that expression of the psbEFLJ operon is a prerequisite for D2 accumulation that is the key regulatory step for D1 accumulation and consecutive assembly of the PSII reaction center complex.

Received for publication, May 24, 2004 , and in revised form, August 20, 2004.

^* This work was supported by Ministry of Education of the Czech Republic, Project LN00A141, Institutional Research Concept AV0Z5020903 (to J. K.), and Deutsche Forschungsgemeinschaft Grants SFB TR1 and SFB 594 (to L. A. E.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^** To whom correspondence should be addressed. Fax: 49-89-17861-209; E-mail: eichacker{at}lmu.de.

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